Heterotetrameric sarcosine oxidase: Structure of a diflavin metalloenzyme at 1.85 A resolution

被引:27
作者
Chen, Zhi-wei
Hassan-Abdulah, Alshaimaa
Zha, Gouhua
Jorns, Marilyn Schuman [1 ]
Mathews, F. Scott
机构
[1] Drexel Univ, Coll Med, Philadelphia, PA 19102 USA
[2] Washington Univ, Sch Med, St Louis, MO 63110 USA
关键词
crystal structure; sarcosine oxidase; flavin mononucleotide; flavin adenine dinucleotide; tetrahydrofolate;
D O I
10.1016/j.jmb.2006.05.067
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
The crystal structure of heterotetrameric sarcosine oxidase (TSOX) from Pseudomonas maltophilia has been determined at 1.85 angstrom resolution. TSOX contains three coenzymes (FAD, FMN and NAD(+)), four different subunits (alpha 103 kDa; beta, 44 kDa; gamma, 21 kDa; delta, 11 kDa) and catalyzes the oxidation of sarcosine (N-methylglycine) to yield hydrogen peroxide, glycine and formaldehyde. In the presence of tetrahydrofolate, the oxidation of sarcosine is coupled to the formation of 5,10-methylenetetrahydrofolate. The NAD(+) and putative folate binding sites are located in the alpha-subunit. The FAD binding site is in the p-subunit. FMN is bound at the interface of the alpha and beta-subunits. The FAD and FMN rings are separated by a short segment of the P-subunit with the closest atoms located 7.4 angstrom apart. Sulfite, an inhibitor of oxygen reduction, is bound at the FMN site. 2-Furoate, a competitive inhibitor with respect to sarcosine, is bound at the FAD site. The sarcosine dehydrogenase and 5,10-methylenetetrahydrofolate synthase sites are 35 angstrom apart but connected by a large internal cavity (similar to 10,000 angstrom(3)). An unexpected zinc ion, coordinated by three cysteine and one histidine side-chains, is bound to the delta-subunit. The N-terminal half of the alpha subunit of TSOX (alpha A) is closely similar to the FAD-binding domain of glutathione reductase but with NAD(+) replacing FAD. The C-terminal half of the alpha subunit of TSOX (alpha B) is similar to the C-terminal half of dimethylglycine oxidase and the T-protein of the glycine cleavage system, proteins that bind tetrahydrofolate. The beta-subunit of TSOX is very similar to monomeric sarcosine oxidase. The gamma-subunit is similar to the C-terminal sub-domain of alpha-TSOX. The 6-subunit shows little similarity with any PDB entry. The alpha A domain/beta-subunit sub-structure of TSOX closely resembles the alpha beta dimer of L-proline dehydrogenase, a heteroctameric protein (alpha beta) that shows highest overall similarity to TSOX. (c) 2006 Elsevier Ltd. All rights reserved.
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页码:1000 / 1018
页数:19
相关论文
共 46 条
[1]   KINETICS OF ELECTRON ENTRY, EXIT, AND INTERFLAVIN ELECTRON-TRANSFER DURING CATALYSIS BY SARCOSINE OXIDASE [J].
ALI, SN ;
ZELLER, HD ;
CALISTO, MK ;
JORNS, MS .
BIOCHEMISTRY, 1991, 30 (45) :10980-10986
[2]   Zinc coordination sphere in biochemical zinc sites [J].
Auld, DS .
BIOMETALS, 2001, 14 (3-4) :271-313
[3]   THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].
BAILEY, S .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763
[4]   FREE R-VALUE - A NOVEL STATISTICAL QUANTITY FOR ASSESSING THE ACCURACY OF CRYSTAL-STRUCTURES [J].
BRUNGER, AT .
NATURE, 1992, 355 (6359) :472-475
[5]  
Brunger AT, 1998, ACTA CRYSTALLOGR D, V54, P905, DOI 10.1107/s0907444998003254
[6]   SEQUENCE-ANALYSIS OF SARCOSINE OXIDASE AND NEARBY GENES REVEALS HOMOLOGIES WITH KEY ENZYMES OF FOLATE ONE-CARBON METABOLISM [J].
CHLUMSKY, LJ ;
ZHANG, LN ;
JORNS, MS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (31) :18252-18259
[7]   PREPARATION AND PROPERTIES OF RECOMBINANT CORYNEBACTERIAL SARCOSINE OXIDASE - EVIDENCE FOR POSTTRANSLATIONAL MODIFICATION DURING TURNOVER WITH SARCOSINE [J].
CHLUMSKY, LJ ;
ZHANG, LN ;
RAMSEY, AJ ;
JORNS, MS .
BIOCHEMISTRY, 1993, 32 (41) :11132-11142
[8]   Identification of the covalent flavin attachment site in sarcosine oxidase [J].
Chlumsky, LJ ;
Sturgess, AW ;
Nieves, E ;
Jorns, MS .
BIOCHEMISTRY, 1998, 37 (08) :2089-2095
[9]   Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods [J].
delaFortelle, E ;
Bricogne, G .
MACROMOLECULAR CRYSTALLOGRAPHY, PT A, 1997, 276 :472-494
[10]   Sequence-structure analysis of FAD-containing proteins [J].
Dym, O ;
Eisenberg, D .
PROTEIN SCIENCE, 2001, 10 (09) :1712-1728