4F21

Crystal structure of carboxylesterase/phospholipase family protein from Francisella tularensis

4F21 の概要

• エントリーDOI: 10.2210/pdb4f21/pdb
• 分子名称: Carboxylesterase/phospholipase family protein, N-((1R,2S)-2-allyl-4-oxocyclobutyl)-4-methylbenzenesulfonamide, bound form (3 entities in total)
• 機能のキーワード: structural genomics, niaid, national institute of allergy and infectious diseases, center for structural genomics of infectious diseases, csgid, carboxylesterase, phospholipase, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• 由来する生物種: Francisella tularensis subsp. tularensis
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 219795.92

構造登録者

Filippova, E.V.,Minasov, G.,Kuhn, M.,Wawrzak, Z.,Shuvalova, L.,Dubrovska, I.,Winsor, J.R.,Kiryukhina, O.,Becker, D.P.,Armoush, N.,Anderson, W.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (登録日: 2012-05-07, 公開日: 2012-08-29, 最終更新日: 2024-10-16)

主引用文献

Filippova, E.V.,Weston, L.A.,Kuhn, M.L.,Geissler, B.,Gehring, A.M.,Armoush, N.,Adkins, C.T.,Minasov, G.,Dubrovska, I.,Shuvalova, L.,Winsor, J.R.,Lavis, L.D.,Satchell, K.J.,Becker, D.P.,Anderson, W.F.,Johnson, R.J.
Large scale structural rearrangement of a serine hydrolase from Francisella tularensis facilitates catalysis.
J.Biol.Chem., 288:10522-10535, 2013

PubMed Abstract: Tularemia is a deadly, febrile disease caused by infection by the gram-negative bacterium, Francisella tularensis. Members of the ubiquitous serine hydrolase protein family are among current targets to treat diverse bacterial infections. Herein we present a structural and functional study of a novel bacterial carboxylesterase (FTT258) from F. tularensis, a homologue of human acyl protein thioesterase (hAPT1). The structure of FTT258 has been determined in multiple forms, and unexpectedly large conformational changes of a peripheral flexible loop occur in the presence of a mechanistic cyclobutanone ligand. The concomitant changes in this hydrophobic loop and the newly exposed hydrophobic substrate binding pocket suggest that the observed structural changes are essential to the biological function and catalytic activity of FTT258. Using diverse substrate libraries, site-directed mutagenesis, and liposome binding assays, we determined the importance of these structural changes to the catalytic activity and membrane binding activity of FTT258. Residues within the newly exposed hydrophobic binding pocket and within the peripheral flexible loop proved essential to the hydrolytic activity of FTT258, indicating that structural rearrangement is required for catalytic activity. Both FTT258 and hAPT1 also showed significant association with liposomes designed to mimic bacterial or human membranes, respectively, even though similar structural rearrangements for hAPT1 have not been reported. The necessity for acyl protein thioesterases to have maximal catalytic activity near the membrane surface suggests that these conformational changes in the protein may dually regulate catalytic activity and membrane association in bacterial and human homologues.

PubMed: 23430251
DOI: 10.1074/jbc.M112.446625

実験手法

X-RAY DIFFRACTION (2.5 Å)