About
Chrystal Starbird completed her undergraduate work at UNC Chapel Hill and spent a few years afterwards working in academic and industry labs before returning to UNC to complete a year-long postbaccalaureate program. Following this, Dr Starbird completed her graduate work in chemical and physical biology at Vanderbilt University. She completed her postdoctoral work as a K99/R00 postdoctoral fellow in the Cancer Biology Center at Yale before moving into her current position as faculty in the Department of Biochemistry and Biophysics at UNC Chapel Hill.
As an expert structural biologist, Dr Starbird runs a research program focused broadly on the molecular basis of disease. Current projects in her lab include studies to improve our understanding of receptor tyrosine kinase activation in diseases such as cancer and the role of lipid transporters in Alzheimer’s disease.
In addition to her research, Dr Starbird is a well-known advocate, mentor, and speaker focused on ways to make science more accessible to all. As a non-traditional student in many ways, Dr Starbird is a strong advocate for inclusive mentoring practices, and publishes regularly on strategies to maximize the impact of mentoring and to build an inclusive lab.
When not in the lab, Chrystal can usually be found searching for the next waterfall to visit with her family (partner and three children).
Selected publications
1485676
{1485676:4IMM4MUP},{1485676:YTUKT2YP},{1485676:LBGAXYWF},{1485676:CJXJLVRT},{1485676:MDDAJKMF},{1485676:XNPSWT6C},{1485676:RG58YANP}
1
modern-language-association
50
date
asc
1
1
1
794
https://biophysicsworkshop.co.za/wp-content/plugins/zotpress/
%7B%22status%22%3A%22success%22%2C%22updateneeded%22%3Afalse%2C%22instance%22%3Afalse%2C%22meta%22%3A%7B%22request_last%22%3A0%2C%22request_next%22%3A0%2C%22used_cache%22%3Atrue%7D%2C%22data%22%3A%5B%7B%22key%22%3A%224IMM4MUP%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Birmingham%20et%20al.%22%2C%22parsedDate%22%3A%222014-05%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EBirmingham%2C%20William%20R.%2C%20et%20al.%20%26%23x201C%3BBioretrosynthetic%20Construction%20of%20a%20Didanosine%20Biosynthetic%20Pathway.%26%23x201D%3B%20%3Ci%3ENature%20Chemical%20Biology%3C%5C%2Fi%3E%2C%20vol.%2010%2C%20no.%205%2C%20May%202014%2C%20pp.%20392%26%23×2013%3B99%2C%20%3Ca%20class%3D%27zp-DOIURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1038%5C%2Fnchembio.1494%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1038%5C%2Fnchembio.1494%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3D4IMM4MUP%27%3ECite%3C%5C%2Fa%3E%20%20%3Ca%20title%3D%27Download%27%20class%3D%27zp-DownloadURL%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.dl.php%3Fapi_user_id%3D1485676%26amp%3Bdlkey%3DMFZFZBTQ%26amp%3Bcontent_type%3Dapplication%5C%2Fpdf%27%3EDownload%3C%5C%2Fa%3E%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Bioretrosynthetic%20construction%20of%20a%20didanosine%20biosynthetic%20pathway%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22William%20R.%22%2C%22lastName%22%3A%22Birmingham%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chrystal%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Timothy%20D.%22%2C%22lastName%22%3A%22Panosian%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22David%20P.%22%2C%22lastName%22%3A%22Nannemann%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22T.%20M.%22%2C%22lastName%22%3A%22Iverson%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Brian%20O.%22%2C%22lastName%22%3A%22Bachmann%22%7D%5D%2C%22abstractNote%22%3A%22Bioretrosynthesis%20is%20meant%20to%20simplify%20construction%20of%20metabolic%20pathways%20by%20screening%20only%20for%20the%20final%20desired%20product.%20This%20approach%2C%20aided%20by%20protein%20design%20and%20crystallography%2C%20is%20now%20used%20to%20synthesize%20an%20antiretroviral%20nucleoside%20analog%20and%20surprisingly%20identifies%20a%20new%20enzyme%20function.%22%2C%22date%22%3A%222014-05%22%2C%22language%22%3A%22en%22%2C%22DOI%22%3A%2210.1038%5C%2Fnchembio.1494%22%2C%22ISSN%22%3A%221552-4469%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.nature.com%5C%2Farticles%5C%2Fnchembio.1494%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A45%3A57Z%22%7D%7D%2C%7B%22key%22%3A%22YTUKT2YP%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Starbird%20et%20al.%22%2C%22parsedDate%22%3A%222015%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EStarbird%2C%20Chrystal%20A.%2C%20et%20al.%20%26%23x201C%3BFlavoenzymes%3A%20Covalent%20versus%20Noncovalent.%26%23x201D%3B%20%3Ci%3EELS%3C%5C%2Fi%3E%2C%20John%20Wiley%20%26amp%3B%20Sons%2C%20Ltd%2C%202015%2C%20pp.%201%26%23×2013%3B11%2C%20https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1002%5C%2F9780470015902.a0026073.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DYTUKT2YP%27%3ECite%3C%5C%2Fa%3E%20%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22bookSection%22%2C%22title%22%3A%22Flavoenzymes%3A%20Covalent%20versus%20Noncovalent%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chrystal%20A%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Elena%22%2C%22lastName%22%3A%22Maklashina%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Gary%22%2C%22lastName%22%3A%22Cecchini%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Tm%22%2C%22lastName%22%3A%22Iverson%22%7D%5D%2C%22abstractNote%22%3A%22The%20use%20of%20nonprotein%20cofactors%20by%20enzymes%20expands%20the%20range%20of%20biological%20chemistries%20supported%20in%20nature.%20Flavins%2C%20which%20are%20derivatives%20of%20vitamin%20B2%2C%20are%20highly%20conjugated%20rings%20that%20are%20particularly%20useful%20for%20oxidoreduction%20and%20group%20transfer%20reactions.%20Most%20flavins%20are%20noncovalently%20associated%20with%20their%20enzymes%2C%20but%20around%2010%25%20of%20flavoproteins%20have%20the%20flavin%20covalently%20attached%20in%20vivo.%20Extensive%20research%20has%20investigated%20how%20the%20presence%20of%20the%20covalent%20bond%20between%20enzyme%20and%20flavin%20cofactor%20influences%20enzymatic%20catalysis.%20This%20work%20identified%20that%20the%20primary%20roles%20of%20the%20covalent%20flavin%20are%20to%20allow%20catalysis%20of%20more%20thermodynamically%20challenging%20reactions%20and%20to%20prevent%20the%20cofactor%20from%20disassociating%20from%20the%20enzyme.%20Major%20questions%20in%20the%20field%20now%20include%20the%20mechanism%20of%20covalent%20flavinylation.%20The%20earliest%20studies%20on%20a%20subset%20of%20covalent%20flavoproteins%20suggested%20that%20cofactor%20attachment%20could%20be%20an%20autocatalytic%20posttranslational%20process.%20However%2C%20the%20recent%20identification%20of%20assembly%20factors%20that%20promote%20covalent%20flavinylation%20identifies%20that%20ancillary%20proteins%20may%20be%20important%20for%20covalent%20flavinylation%20in%20vivo.%20Key%20Concepts%20Covalent%20flavin%20attachment%20increases%20stability%20of%20the%20holoenzyme%20and%20increases%20the%20enzyme%27s%20redox%20potential.%20Covalent%20flavinylation%20may%20occur%20either%20through%20an%20entirely%20autocatalytic%20mechanism%2C%20or%20be%20assisted%20by%20assembly%20factors.%20Enzyme-associated%20flavin%20can%20promote%20a%20variety%20of%20chemistries.%20Flavoenzymes%20can%20have%20covalent%20or%20noncovalent%20flavin.%20Covalent%20flavinylation%20can%20occur%20on%20multiple%20sites%20of%20the%20protein%20and%20flavin%20molecule.%22%2C%22bookTitle%22%3A%22eLS%22%2C%22date%22%3A%222015%22%2C%22language%22%3A%22en%22%2C%22ISBN%22%3A%22978-0-470-01590-2%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fonlinelibrary.wiley.com%5C%2Fdoi%5C%2Fabs%5C%2F10.1002%5C%2F9780470015902.a0026073%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A45%3A22Z%22%7D%7D%2C%7B%22key%22%3A%22LBGAXYWF%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Immormino%20et%20al.%22%2C%22parsedDate%22%3A%222015-06-09%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EImmormino%2C%20Robert%20M.%2C%20et%20al.%20%26%23x201C%3BProbing%20Mechanistic%20Similarities%20between%20Response%20Regulator%20Signaling%20Proteins%20and%20Haloacid%20Dehalogenase%20Phosphatases.%26%23x201D%3B%20%3Ci%3EBiochemistry%3C%5C%2Fi%3E%2C%20vol.%2054%2C%20no.%2022%2C%20June%202015%2C%20pp.%203514%26%23×2013%3B27%2C%20%3Ca%20class%3D%27zp-ItemURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.5b00286%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.5b00286%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DLBGAXYWF%27%3ECite%3C%5C%2Fa%3E%20%20%3Ca%20title%3D%27Download%27%20class%3D%27zp-DownloadURL%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.dl.php%3Fapi_user_id%3D1485676%26amp%3Bdlkey%3D956Q3GLD%26amp%3Bcontent_type%3Dapplication%5C%2Fpdf%27%3EDownload%3C%5C%2Fa%3E%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Probing%20Mechanistic%20Similarities%20between%20Response%20Regulator%20Signaling%20Proteins%20and%20Haloacid%20Dehalogenase%20Phosphatases%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Robert%20M.%22%2C%22lastName%22%3A%22Immormino%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chrystal%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Ruth%20E.%22%2C%22lastName%22%3A%22Silversmith%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Robert%20B.%22%2C%22lastName%22%3A%22Bourret%22%7D%5D%2C%22abstractNote%22%3A%22Response%20regulator%20signaling%20proteins%20and%20phosphatases%20of%20the%20haloacid%20dehalogenase%20%28HAD%29%20superfamily%20share%20strikingly%20similar%20folds%2C%20active%20site%20geometries%2C%20and%20reaction%20chemistry.%20Proteins%20from%20both%20families%20catalyze%20the%20transfer%20of%20a%20phosphoryl%20group%20from%20a%20substrate%20to%20one%20of%20their%20own%20aspartyl%20residues%2C%20and%20subsequent%20hydrolysis%20of%20the%20phosphoprotein.%20Notable%20differences%20include%20an%20additional%20Asp%20that%20functions%20as%20an%20acid%5C%2Fbase%20catalyst%20and%20an%20active%20site%20well-structured%20prior%20to%20phosphorylation%20in%20HAD%20phosphatases.%20Both%20features%20contribute%20to%20reactions%20substantially%20faster%20than%20those%20for%20response%20regulators.%20To%20investigate%20mechanisms%20underlying%20the%20functional%20differences%20between%20response%20regulators%20and%20HAD%20phosphatases%2C%20we%20characterized%20five%20double%20mutants%20of%20the%20response%20regulator%20CheY%20designed%20to%20mimic%20HAD%20phosphatases.%20Each%20mutant%20contained%20the%20extra%20Asp%20paired%20with%20a%20phosphatase-inspired%20substitution%20to%20potentially%20position%20the%20Asp%20properly.%20Only%20CheY%20DR%20%28Arg%20as%20the%20anchor%29%20exhibited%20enhanced%20rates%20of%20both%20autophosphorylation%20with%20phosphoramidate%20and%20autodephosphorylation%20compared%20to%20those%20of%20wild-type%20CheY.%20Crystal%20structures%20of%20CheY%20DR%20complexed%20with%20MoO42%5Cu2013%20or%20WO42%5Cu2013%20revealed%20active%20site%20hydrogen%20bonding%20networks%20similar%20to%20those%20in%20HAD%5Cu00b7substrate%20complexes%2C%20with%20the%20extra%20Asp%20positioned%20for%20direct%20interaction%20with%20the%20leaving%20group%20%28phosphorylation%29%20or%20nucleophile%20%28dephosphorylation%29.%20However%2C%20CheY%20DR%20reaction%20kinetics%20did%20not%20exhibit%20the%20pH%20sensitivities%20expected%20for%20acid%5C%2Fbase%20catalysis.%20Biochemical%20analysis%20indicated%20CheY%20DR%20had%20an%20enhanced%20propensity%20to%20adopt%20the%20active%20conformation%20without%20phosphorylation%2C%20but%20a%20crystal%20structure%20revealed%20unphosphorylated%20CheY%20DR%20was%20not%20locked%20in%20the%20active%20conformation.%20Thus%2C%20the%20enhanced%20reactivity%20of%20CheY%20DR%20reflected%20partial%20acquisition%20of%20catalytic%20and%20structural%20features%20of%20HAD%20phosphatases.%22%2C%22date%22%3A%222015-06-09%22%2C%22language%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Facs.biochem.5b00286%22%2C%22ISSN%22%3A%220006-2960%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.5b00286%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A43%3A56Z%22%7D%7D%2C%7B%22key%22%3A%22CJXJLVRT%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Maklashina%20et%20al.%22%2C%22parsedDate%22%3A%222016-02-01%22%2C%22numChildren%22%3A2%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EMaklashina%2C%20Elena%2C%20et%20al.%20%26%23x201C%3BBinding%20of%20the%20Covalent%20Flavin%20Assembly%20Factor%20to%20the%20Flavoprotein%20Subunit%20of%20Complex%20II%20%2A.%26%23x201D%3B%20%3Ci%3EJournal%20of%20Biological%20Chemistry%3C%5C%2Fi%3E%2C%20vol.%20291%2C%20no.%206%2C%20Feb.%202016%2C%20pp.%202904%26%23×2013%3B16%2C%20%3Ca%20class%3D%27zp-DOIURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1074%5C%2Fjbc.M115.690396%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1074%5C%2Fjbc.M115.690396%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DCJXJLVRT%27%3ECite%3C%5C%2Fa%3E%20%20%3Ca%20title%3D%27Download%27%20class%3D%27zp-DownloadURL%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.dl.php%3Fapi_user_id%3D1485676%26amp%3Bdlkey%3D2PB789VK%26amp%3Bcontent_type%3Dapplication%5C%2Fpdf%27%3EDownload%3C%5C%2Fa%3E%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Binding%20of%20the%20Covalent%20Flavin%20Assembly%20Factor%20to%20the%20Flavoprotein%20Subunit%20of%20Complex%20II%20%2A%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Elena%22%2C%22lastName%22%3A%22Maklashina%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Sany%22%2C%22lastName%22%3A%22Rajagukguk%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chrystal%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22W.%20Hayes%22%2C%22lastName%22%3A%22McDonald%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Anna%22%2C%22lastName%22%3A%22Koganitsky%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Michael%22%2C%22lastName%22%3A%22Eisenbach%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Tina%20M.%22%2C%22lastName%22%3A%22Iverson%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Gary%22%2C%22lastName%22%3A%22Cecchini%22%7D%5D%2C%22abstractNote%22%3A%22%3Cp%3E%3Ci%3EEscherichia%20coli%3C%5C%2Fi%3E%20harbors%20two%20highly%20conserved%20homologs%20of%20the%20essential%20mitochondrial%20respiratory%20complex%20II%20%28succinate%3Aubiquinone%20oxidoreductase%29.%20Aerobically%20the%20bacterium%20synthesizes%20succinate%3Aquinone%20reductase%20as%20part%20of%20its%20respiratory%20chain%2C%20whereas%20under%20microaerophilic%20conditions%2C%20the%20quinol%3Afumarate%20reductase%20can%20be%20utilized.%20All%20complex%20II%20enzymes%20harbor%20a%20covalently%20bound%20FAD%20co-factor%20that%20is%20essential%20for%20their%20ability%20to%20oxidize%20succinate.%20In%20eukaryotes%20and%20many%20bacteria%2C%20assembly%20of%20the%20covalent%20flavin%20linkage%20is%20facilitated%20by%20a%20small%20protein%20assembly%20factor%2C%20termed%20SdhE%20in%20%3Ci%3EE.%20coli%3C%5C%2Fi%3E.%20How%20SdhE%20assists%20with%20formation%20of%20the%20covalent%20flavin%20bond%20and%20how%20it%20binds%20the%20flavoprotein%20subunit%20of%20complex%20II%20remain%20unknown.%20Using%20photo-cross-linking%2C%20we%20report%20the%20interaction%20site%20between%20the%20flavoprotein%20of%20complex%20II%20and%20the%20SdhE%20assembly%20factor.%20These%20data%20indicate%20that%20SdhE%20binds%20to%20the%20flavoprotein%20between%20two%20independently%20folded%20domains%20and%20that%20this%20binding%20mode%20likely%20influences%20the%20interdomain%20orientation.%20In%20so%20doing%2C%20SdhE%20likely%20orients%20amino%20acid%20residues%20near%20the%20dicarboxylate%20and%20FAD%20binding%20site%2C%20which%20facilitates%20formation%20of%20the%20covalent%20flavin%20linkage.%20These%20studies%20identify%20how%20the%20conserved%20SdhE%20assembly%20factor%20and%20its%20homologs%20participate%20in%20complex%20II%20maturation.%3C%5C%2Fp%3E%22%2C%22date%22%3A%222016%5C%2F02%5C%2F01%22%2C%22language%22%3A%22English%22%2C%22DOI%22%3A%2210.1074%5C%2Fjbc.M115.690396%22%2C%22ISSN%22%3A%220021-9258%2C%201083-351X%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.jbc.org%5C%2Farticle%5C%2FS0021-9258%2820%2934278-2%5C%2Fabstract%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A43%3A14Z%22%7D%7D%2C%7B%22key%22%3A%22MDDAJKMF%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Starbird%20et%20al.%22%2C%22parsedDate%22%3A%222017-08-04%22%2C%22numChildren%22%3A2%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EStarbird%2C%20C.%20A.%2C%20et%20al.%20%26%23x201C%3BStructural%20and%20Biochemical%20Analyses%20Reveal%20Insights%20into%20Covalent%20Flavinylation%20of%20the%20Escherichia%20Coli%20Complex%20II%20Homolog%20Quinol%3AFumarate%20Reductase.%26%23x201D%3B%20%3Ci%3EJournal%20of%20Biological%20Chemistry%3C%5C%2Fi%3E%2C%20vol.%20292%2C%20no.%2031%2C%20Aug.%202017%2C%20pp.%2012921%26%23×2013%3B33%2C%20%3Ca%20class%3D%27zp-DOIURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1074%5C%2Fjbc.M117.795120%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1074%5C%2Fjbc.M117.795120%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DMDDAJKMF%27%3ECite%3C%5C%2Fa%3E%20%20%3Ca%20title%3D%27Download%27%20class%3D%27zp-DownloadURL%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.dl.php%3Fapi_user_id%3D1485676%26amp%3Bdlkey%3DCWIQVGJE%26amp%3Bcontent_type%3Dapplication%5C%2Fpdf%27%3EDownload%3C%5C%2Fa%3E%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Structural%20and%20biochemical%20analyses%20reveal%20insights%20into%20covalent%20flavinylation%20of%20the%20Escherichia%20coli%20Complex%20II%20homolog%20quinol%3Afumarate%20reductase%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22C.%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Elena%22%2C%22lastName%22%3A%22Maklashina%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Pankaj%22%2C%22lastName%22%3A%22Sharma%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Susan%22%2C%22lastName%22%3A%22Qualls-Histed%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Gary%22%2C%22lastName%22%3A%22Cecchini%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22T.%20M.%22%2C%22lastName%22%3A%22Iverson%22%7D%5D%2C%22abstractNote%22%3A%22%3Cp%3EThe%20%3Ci%3EEscherichia%20coli%3C%5C%2Fi%3E%20Complex%20II%20homolog%20quinol%3Afumarate%20reductase%20%28QFR%2C%20FrdABCD%29%20catalyzes%20the%20interconversion%20of%20fumarate%20and%20succinate%20at%20a%20covalently%20attached%20FAD%20within%20the%20FrdA%20subunit.%20The%20SdhE%20assembly%20factor%20enhances%20covalent%20flavinylation%20of%20Complex%20II%20homologs%2C%20but%20the%20mechanisms%20underlying%20the%20covalent%20attachment%20of%20FAD%20remain%20to%20be%20fully%20elucidated.%20Here%2C%20we%20explored%20the%20mechanisms%20of%20covalent%20flavinylation%20of%20the%20%3Ci%3EE.%20coli%3C%5C%2Fi%3E%20QFR%20FrdA%20subunit.%20Using%20a%20%5Cu0394%3Ci%3EsdhE%20E.%20coli%3C%5C%2Fi%3E%20strain%2C%20we%20show%20that%20the%20requirement%20for%20the%20assembly%20factor%20depends%20on%20the%20cellular%20redox%20environment.%20We%20next%20identified%20residues%20important%20for%20the%20covalent%20attachment%20and%20selected%20the%20FrdA%3Csup%3EE245%3C%5C%2Fsup%3E%20residue%2C%20which%20contributes%20to%20proton%20shuttling%20during%20fumarate%20reduction%2C%20for%20detailed%20biophysical%20and%20structural%20characterization.%20We%20found%20that%20QFR%20complexes%20containing%20FrdA%3Csup%3EE245Q%3C%5C%2Fsup%3E%20have%20a%20structure%20similar%20to%20that%20of%20the%20WT%20flavoprotein%2C%20but%20lack%20detectable%20substrate%20binding%20and%20turnover.%20In%20the%20context%20of%20the%20isolated%20FrdA%20subunit%2C%20the%20anticipated%20assembly%20intermediate%20during%20covalent%20flavinylation%2C%20FrdA%3Csup%3EE245%3C%5C%2Fsup%3E%20variants%20had%20stability%20similar%20to%20that%20of%20WT%20FrdA%2C%20contained%20noncovalent%20FAD%2C%20and%20displayed%20a%20reduced%20capacity%20to%20interact%20with%20SdhE.%20However%2C%20small-angle%20X-ray%20scattering%20%28SAXS%29%20analysis%20of%20WT%20FrdA%20cross-linked%20to%20SdhE%20suggested%20that%20the%20FrdA%3Csup%3EE245%3C%5C%2Fsup%3E%20residue%20is%20unlikely%20to%20contribute%20directly%20to%20the%20FrdA-SdhE%20protein-protein%20interface.%20We%20also%20found%20that%20no%20auxiliary%20factor%20is%20absolutely%20required%20for%20flavinylation%2C%20indicating%20that%20the%20covalent%20flavinylation%20is%20autocatalytic.%20We%20propose%20that%20multiple%20factors%2C%20including%20the%20SdhE%20assembly%20factor%20and%20bound%20dicarboxylates%2C%20stimulate%20covalent%20flavinylation%20by%20preorganizing%20the%20active%20site%20to%20stabilize%20the%20quinone-methide%20intermediate.%3C%5C%2Fp%3E%22%2C%22date%22%3A%222017%5C%2F08%5C%2F04%22%2C%22language%22%3A%22English%22%2C%22DOI%22%3A%2210.1074%5C%2Fjbc.M117.795120%22%2C%22ISSN%22%3A%220021-9258%2C%201083-351X%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.jbc.org%5C%2Farticle%5C%2FS0021-9258%2820%2940063-8%5C%2Fabstract%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A42%3A34Z%22%7D%7D%2C%7B%22key%22%3A%22XNPSWT6C%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Starbird%20et%20al.%22%2C%22parsedDate%22%3A%222018-12-18%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EStarbird%2C%20C.%20A.%2C%20et%20al.%20%26%23x201C%3BThe%20Structure%20of%20the%20Bifunctional%20Everninomicin%20Biosynthetic%20Enzyme%20EvdMO1%20Suggests%20Independent%20Activity%20of%20the%20Fused%20Methyltransferase-Oxidase%20Domains.%26%23x201D%3B%20%3Ci%3EBiochemistry%3C%5C%2Fi%3E%2C%20vol.%2057%2C%20no.%2050%2C%20Dec.%202018%2C%20pp.%206827%26%23×2013%3B37%2C%20%3Ca%20class%3D%27zp-ItemURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.8b00836%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.8b00836%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DXNPSWT6C%27%3ECite%3C%5C%2Fa%3E%20%20%3Ca%20title%3D%27Download%27%20class%3D%27zp-DownloadURL%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.dl.php%3Fapi_user_id%3D1485676%26amp%3Bdlkey%3DM6CIG9CQ%26amp%3Bcontent_type%3Dapplication%5C%2Fpdf%27%3EDownload%3C%5C%2Fa%3E%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22The%20Structure%20of%20the%20Bifunctional%20Everninomicin%20Biosynthetic%20Enzyme%20EvdMO1%20Suggests%20Independent%20Activity%20of%20the%20Fused%20Methyltransferase-Oxidase%20Domains%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22C.%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Nicole%20A.%22%2C%22lastName%22%3A%22Perry%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Qiuyan%22%2C%22lastName%22%3A%22Chen%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Sandra%22%2C%22lastName%22%3A%22Berndt%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Izumi%22%2C%22lastName%22%3A%22Yamakawa%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Lioudmila%20V.%22%2C%22lastName%22%3A%22Loukachevitch%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Emilianne%20M.%22%2C%22lastName%22%3A%22Limbrick%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Brian%20O.%22%2C%22lastName%22%3A%22Bachmann%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22T.%20M.%22%2C%22lastName%22%3A%22Iverson%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kathryn%20M.%22%2C%22lastName%22%3A%22McCulloch%22%7D%5D%2C%22abstractNote%22%3A%22Members%20of%20the%20orthosomycin%20family%20of%20natural%20products%20are%20decorated%20polysaccharides%20with%20potent%20antibiotic%20activity%20and%20complex%20biosynthetic%20pathways.%20The%20defining%20feature%20of%20the%20orthosomycins%20is%20an%20orthoester%20linkage%20between%20carbohydrate%20moieties%20that%20is%20necessary%20for%20antibiotic%20activity%20and%20is%20likely%20formed%20by%20a%20family%20of%20conserved%20oxygenases.%20Everninomicins%20are%20octasaccharide%20orthosomycins%20produced%20by%20Micromonospora%20carbonacea%20that%20have%20two%20orthoester%20linkages%20and%20a%20methylenedioxy%20bridge%2C%20three%20features%20whose%20formation%20logically%20requires%20oxidative%20chemistry.%20Correspondingly%2C%20the%20evd%20gene%20cluster%20encoding%20everninomicin%20D%20encodes%20two%20monofunctional%20nonheme%20iron%2C%20%5Cu03b1-ketoglutarate-dependent%20oxygenases%20and%20one%20bifunctional%20enzyme%20with%20an%20N-terminal%20methyltransferase%20domain%20and%20a%20C-terminal%20oxygenase%20domain.%20To%20investigate%20whether%20the%20activities%20of%20these%20domains%20are%20linked%20in%20the%20bifunctional%20enzyme%20EvdMO1%2C%20we%20determined%20the%20structure%20of%20the%20N-terminal%20methyltransferase%20domain%20to%201.1%20%5Cu00c5%20and%20that%20of%20the%20full-length%20protein%20to%203.35%20%5Cu00c5%20resolution.%20Both%20domains%20of%20EvdMO1%20adopt%20the%20canonical%20folds%20of%20their%20respective%20superfamilies%20and%20are%20connected%20by%20a%20short%20linker.%20Each%20domain%5Cu2019s%20active%20site%20is%20oriented%20such%20that%20it%20faces%20away%20from%20the%20other%20domain%2C%20and%20there%20is%20no%20evidence%20of%20a%20channel%20connecting%20the%20two.%20Our%20results%20support%20EvdMO1%20working%20as%20a%20bifunctional%20enzyme%20with%20independent%20catalytic%20activities.%22%2C%22date%22%3A%222018-12-18%22%2C%22language%22%3A%22%22%2C%22DOI%22%3A%2210.1021%5C%2Facs.biochem.8b00836%22%2C%22ISSN%22%3A%220006-2960%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1021%5C%2Facs.biochem.8b00836%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A41%3A10Z%22%7D%7D%2C%7B%22key%22%3A%22RG58YANP%22%2C%22library%22%3A%7B%22id%22%3A1485676%7D%2C%22meta%22%3A%7B%22creatorSummary%22%3A%22Bagchi%20et%20al.%22%2C%22parsedDate%22%3A%222024-09-05%22%2C%22numChildren%22%3A1%7D%2C%22bib%22%3A%22%3Cdiv%20class%3D%5C%22csl-bib-body%5C%22%20style%3D%5C%22line-height%3A%202%3B%20padding-left%3A%201em%3B%20text-indent%3A-1em%3B%5C%22%3E%5Cn%20%20%3Cdiv%20class%3D%5C%22csl-entry%5C%22%3EBagchi%2C%20Atrish%2C%20et%20al.%20%26%23x201C%3BStructural%20Insights%20into%20the%20Role%20and%20Targeting%20of%20EGFRvIII.%26%23x201D%3B%20%3Ci%3EStructure%3C%5C%2Fi%3E%2C%20vol.%2032%2C%20no.%209%2C%20Sept.%202024%2C%20pp.%201367-1380.e6%2C%20%3Ca%20class%3D%27zp-DOIURL%27%20href%3D%27https%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1016%5C%2Fj.str.2024.05.018%27%3Ehttps%3A%5C%2F%5C%2Fdoi.org%5C%2F10.1016%5C%2Fj.str.2024.05.018%3C%5C%2Fa%3E.%20%3Ca%20title%3D%27Cite%20in%20RIS%20Format%27%20class%3D%27zp-CiteRIS%27%20href%3D%27https%3A%5C%2F%5C%2Fbiophysicsworkshop.co.za%5C%2Fwp-content%5C%2Fplugins%5C%2Fzotpress%5C%2Flib%5C%2Frequest%5C%2Frequest.cite.php%3Fapi_user_id%3D1485676%26amp%3Bitem_key%3DRG58YANP%27%3ECite%3C%5C%2Fa%3E%20%3C%5C%2Fdiv%3E%5Cn%3C%5C%2Fdiv%3E%22%2C%22data%22%3A%7B%22itemType%22%3A%22journalArticle%22%2C%22title%22%3A%22Structural%20insights%20into%20the%20role%20and%20targeting%20of%20EGFRvIII%22%2C%22creators%22%3A%5B%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Atrish%22%2C%22lastName%22%3A%22Bagchi%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Steven%20E.%22%2C%22lastName%22%3A%22Stayrook%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Katerina%20T.%22%2C%22lastName%22%3A%22Xenaki%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Chrystal%20A.%22%2C%22lastName%22%3A%22Starbird%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Sofia%22%2C%22lastName%22%3A%22Doulkeridou%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Rachid%20el%22%2C%22lastName%22%3A%22Khoulati%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Rob%20C.%22%2C%22lastName%22%3A%22Roovers%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Karl%20R.%22%2C%22lastName%22%3A%22Schmitz%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Paul%20M.%20P.%20van%20Bergen%20en%22%2C%22lastName%22%3A%22Henegouwen%22%7D%2C%7B%22creatorType%22%3A%22author%22%2C%22firstName%22%3A%22Kathryn%20M.%22%2C%22lastName%22%3A%22Ferguson%22%7D%5D%2C%22abstractNote%22%3A%22%22%2C%22date%22%3A%222024-09-05%22%2C%22language%22%3A%22English%22%2C%22DOI%22%3A%2210.1016%5C%2Fj.str.2024.05.018%22%2C%22ISSN%22%3A%220969-2126%22%2C%22url%22%3A%22https%3A%5C%2F%5C%2Fwww.cell.com%5C%2Fstructure%5C%2Fabstract%5C%2FS0969-2126%2824%2900195-3%22%2C%22collections%22%3A%5B%22EENXQ3XN%22%5D%2C%22dateModified%22%3A%222025-05-17T14%3A40%3A07Z%22%7D%7D%5D%7D
Birmingham, William R., et al. “Bioretrosynthetic Construction of a Didanosine Biosynthetic Pathway.”
Nature Chemical Biology, vol. 10, no. 5, May 2014, pp. 392–99,
https://doi.org/10.1038/nchembio.1494.
Cite Download
Starbird, Chrystal A., et al. “Flavoenzymes: Covalent versus Noncovalent.”
ELS, John Wiley & Sons, Ltd, 2015, pp. 1–11, https://doi.org/10.1002/9780470015902.a0026073.
Cite
Immormino, Robert M., et al. “Probing Mechanistic Similarities between Response Regulator Signaling Proteins and Haloacid Dehalogenase Phosphatases.”
Biochemistry, vol. 54, no. 22, June 2015, pp. 3514–27,
https://doi.org/10.1021/acs.biochem.5b00286.
Cite Download
Maklashina, Elena, et al. “Binding of the Covalent Flavin Assembly Factor to the Flavoprotein Subunit of Complex II *.”
Journal of Biological Chemistry, vol. 291, no. 6, Feb. 2016, pp. 2904–16,
https://doi.org/10.1074/jbc.M115.690396.
Cite Download
Starbird, C. A., et al. “Structural and Biochemical Analyses Reveal Insights into Covalent Flavinylation of the Escherichia Coli Complex II Homolog Quinol:Fumarate Reductase.”
Journal of Biological Chemistry, vol. 292, no. 31, Aug. 2017, pp. 12921–33,
https://doi.org/10.1074/jbc.M117.795120.
Cite Download
Starbird, C. A., et al. “The Structure of the Bifunctional Everninomicin Biosynthetic Enzyme EvdMO1 Suggests Independent Activity of the Fused Methyltransferase-Oxidase Domains.”
Biochemistry, vol. 57, no. 50, Dec. 2018, pp. 6827–37,
https://doi.org/10.1021/acs.biochem.8b00836.
Cite Download
Bagchi, Atrish, et al. “Structural Insights into the Role and Targeting of EGFRvIII.”
Structure, vol. 32, no. 9, Sept. 2024, pp. 1367-1380.e6,
https://doi.org/10.1016/j.str.2024.05.018.
Cite