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Determination of unknown protein function using crystallographic screening of metabolite cocktail binding
Author(s) -
Shumilin Igor A.,
Cymborowski Marcin,
Chertihin Olga,
Jha Kula N.,
Herr John C.,
Lesley Scott A.,
Joachimiak Andrzej,
Minor Wladek
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb107
Subject(s) - biochemistry , metabolite , archaea , function (biology) , bacillus subtilis , biology , protein family , computational biology , gene , chemistry , genetics , bacteria
An estimated 30–40% of sequenced bacterial genes, and an even higher fraction of genes in archaea and eukaryotes, encode for proteins that lack an assigned function. Identification of unknown protein functions is a high‐priority task in our efforts to understand cellular processes. Here, we applied crystallographic screening to assess the binding of compounds in a metabolite library to proteins in two families. These two families are the YjeF_N family of unknown function represented by a mouse apolipoprotein‐I binding protein and the N‐terminal domain of Tm0922 from Thermotoga maritima , and the PF01256 family, previously annotated as a family of small molecule kinases, represented by YxkO from Bacillus subtilis and the C‐terminal domain of Tm0922. The selective binding of several structurally‐related compounds observed for each protein provides the basis for identification of the likely ligands of the proteins. Subsequently, the PF01256 proteins are shown to catalyze ATP‐dependent NAD(P)H‐hydrate dehydration, the previously described orphan activity. The YjeF_N proteins interact with an adenosine diphosphoribose‐related (ADPR) substrate and most likely serve as ADPR transferases. Metabolite library screening may be used as an efficient tool for the functional studies of uncharacterized proteins. The funding was provided by the NIH grants U01‐HD060491, U54‐GM74492, U54‐GM074942 and R01‐GM53163.