8SUT
Crystal structure of YisK from Bacillus subtilis in complex with reaction product 4-Hydroxy-2-oxoglutaric acid
8SUT の概要
エントリーDOI | 10.2210/pdb8sut/pdb |
関連するPDBエントリー | 8SKY |
分子名称 | Fumarylacetoacetate hydrolase family protein, (2~{R})-2-oxidanyl-4-oxidanylidene-pentanedioic acid, MANGANESE (II) ION, ... (4 entities in total) |
機能のキーワード | fumarylacetoacetate hydrolase, decarboxylase, elongasome, hydrolase |
由来する生物種 | Bacillus subtilis |
タンパク質・核酸の鎖数 | 2 |
化学式量合計 | 69534.95 |
構造登録者 | Krieger, I.V.,Chemelewski, V.,Guo, T.,Sperber, A.,Herman, J.,Sacchettini, J.C. (登録日: 2023-05-13, 公開日: 2023-11-01, 最終更新日: 2024-02-07) |
主引用文献 | Guo, T.,Sperber, A.M.,Krieger, I.V.,Duan, Y.,Chemelewski, V.R.,Sacchettini, J.C.,Herman, J.K. Bacillus subtilis YisK possesses oxaloacetate decarboxylase activity and exhibits Mbl-dependent localization. J.Bacteriol., 206:e0020223-e0020223, 2024 Cited by PubMed Abstract: YisK is an uncharacterized protein in previously shown to interact genetically with the elongasome protein Mbl. YisK overexpression leads to cell widening and lysis, phenotypes that are dependent on and suppressed by mutations. In the present work, we characterize YisK's localization, structure, and enzymatic activity. We show that YisK localizes as puncta that depend on Mbl. YisK belongs to the fumarylacetoacetate hydrolase (FAH) superfamily, and crystal structures revealed close structural similarity to two oxaloacetate (OAA) decarboxylases: human mitochondrial FAHD1 and Cg1458. We demonstrate that YisK can also catalyze the decarboxylation of OAA ( = 134 µM, = 31 min). A catalytic dead variant (YisK E148A, E150A) retains wild-type localization and still widens cells following overexpression, indicating these activities are not dependent on YisK catalysis. Conversely, a non-localizing variant (YisK E30A) retains wild-type enzymatic activity but localizes diffusely and no longer widens cells following overexpression. Together, these results suggest that YisK may be subject to spatial regulation that depends on the cell envelope synthesis machinery. IMPORTANCE The elongasome is a multiprotein complex that guides lengthwise growth in some bacteria. We previously showed that, in , overexpression of an uncharacterized putative enzyme (YisK) perturbed function of the actin-like elongasome protein Mbl. Here, we show that YisK exhibits Mbl-dependent localization. Through biochemical and structural characterization, we demonstrate that, like its mitochondrial homolog FAHD1, YisK can catalyze the decarboxylation of the oxaloacetate to pyruvate and CO. YisK is the first example of an enzyme implicated in central carbon metabolism with subcellular localization that depends on Mbl. PubMed: 38047707DOI: 10.1128/jb.00202-23 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (1.93 Å) |
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