Crystal structure and fluorescence studies reveal the role of helical dimeric interface of staphylococcal fabg1 in positive cooperativity for NADPH

被引：23

作者：

Dutta, Debajyoti ^{[1
]}

Bhattacharyya, Sudipta ^{[1
]}

Das, Amit Kumar ^{[1
]}

机构：

[1] Indian Inst Technol, Dept Biotechnol, Kharagpur 721302, W Bengal, India

来源：

PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS | 2012年 / 80卷 / 04期

关键词：

dimerization axis; fluorescence resonance energy transfer; hypsochromic shift; positive cooperativity; Staphylococcus aureus; tryptophan fluorescence; PLASMODIUM-FALCIPARUM; REDUCTASE; COFACTOR; PATHWAY;

D O I：

10.1002/prot.24024

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Crystal structure of Staphylococcal beta-ketoacyl-ACP reductase 1 (SaFabG1) complexed with NADPH is determined at 2.5 angstrom resolution. The enzyme is essential in FAS-II pathway and utilizes NADPH to reduce beta-ketoacyl-ACP to (S)-beta-hydroxyacyl-ACP. Unlike the tetrameric FabGs, dimeric SaFabG1 shows positive homotropic cooperativity towards NADPH. Analysis of FabG:NADPH binary crystal structure endorses that NADPH interacts directly with the helices a4 and a5 those are present on a dimerization interface. A steady shift in tryptophan (of a4 helix) emission peak upon steady increment of NADPH concentration reveals that the dimeric interface is formed by a4-a4' and a5-a5' helices. This dimeric interface imparts positive homotropic cooperativity towards NADPH. PEG, a substrate mimicking molecule is also found near the active site of the enzyme. Proteins 2012; (c) 2011 Wiley Periodicals, Inc.

引用

页码：1250 / 1257

页数：8

共 24 条

[1] THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].

BAILEY, S .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763

[2] Essentiality of FASII pathway for Staphylococcus aureus [J].

Balemans, Wendy ;

Lounis, Nacer ;

Gilissen, Ron ;

Guillemont, Jerome ;

Simmen, Kenny ;

Andries, Koen ;

Koul, Anil .

NATURE, 2010, 463 (7279) :E3-E4

[3] Crystal structure of FabG4 from Mycobacterium tuberculosis reveals the importance of C-terminal residues in ketoreductase activity [J].

Dutta, Debajyoti ;

Bhattacharyya, Sudipta ;

Mukherjee, Somnath ;

Saha, Baisakhee ;

Das, Amit Kumar .

JOURNAL OF STRUCTURAL BIOLOGY, 2011, 174 (01) :147-155

[4] Coot:: model-building tools for molecular graphics [J].

Emsley, P ;

Cowtan, K .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2004, 60 :2126-2132

[5]

Evans P.R., 1993, Proceedings of the CCP4 Study Weekend. Data Collection and Processing, P114

[6] SHORT-CHAIN DEHYDROGENASES REDUCTASES (SDR) [J].

JORNVALL, H ;

PERSSON, B ;

KROOK, M ;

ATRIAN, S ;

GONZALEZDUARTE, R ;

JEFFERY, J ;

GHOSH, D .

BIOCHEMISTRY, 1995, 34 (18) :6003-6013

[7] AUTOMATIC PROCESSING OF ROTATION DIFFRACTION DATA FROM CRYSTALS OF INITIALLY UNKNOWN SYMMETRY AND CELL CONSTANTS [J].

KABSCH, W .

JOURNAL OF APPLIED CRYSTALLOGRAPHY, 1993, 26 :795-800

[8] Analyses of co-operative transitions in Plasmodium falciparum β-ketoacyl acyl carrier protein reductase upon co-factor and acyl carrier protein binding [J].

Karmodiya, Krishanpal ;

Surolia, Namita .

FEBS JOURNAL, 2006, 273 (17) :4093-4103

[9] Inference of macromolecular assemblies from crystalline state [J].

Krissinel, Evgeny ;

Henrick, Kim .

JOURNAL OF MOLECULAR BIOLOGY, 2007, 372 (03) :774-797

[10]

Lakowicz J.R., 2004, PRINCIPLES FLUORESCE, V2nd

← 1 2 3 →