4WKC

Crystal structure of Escherichia coli 5'-methylthioadenosine/S-adenosyl homocysteine nucleosidase (MTAN) complexed with butylthio-DADMe-Immucillin-A

Summary for 4WKC

• Entry DOI: 10.2210/pdb4wkc/pdb
• Related: 1Y6Q 4WKN 4WKO 4WKP
• Descriptor: 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase, (3R,4S)-1-[(4-amino-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl]-4-[(butylsulfanyl)methyl]pyrrolidin-3-ol, TETRAETHYLENE GLYCOL, ... (4 entities in total)
• Functional Keywords: hydrolase, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Escherichia coli O139:H28
• Total number of polymer chains: 1
• Total formula weight: 26588.40

Authors

Cameron, S.A.,Thomas, K.,Almo, S.C.,Schramm, V.L. (deposition date: 2014-10-02, release date: 2015-08-19, Last modification date: 2023-09-27)

Primary citation

Thomas, K.,Cameron, S.A.,Almo, S.C.,Burgos, E.S.,Gulab, S.A.,Schramm, V.L.
Active site and remote contributions to catalysis in methylthioadenosine nucleosidases.
Biochemistry, 54:2520-2529, 2015

PubMed Abstract: 5'-Methylthioadenosine/S-adenosyl-l-homocysteine nucleosidases (MTANs) catalyze the hydrolysis of 5'-methylthioadenosine to adenine and 5-methylthioribose. The amino acid sequences of the MTANs from Vibrio cholerae (VcMTAN) and Escherichia coli (EcMTAN) are 60% identical and 75% similar. Protein structure folds and kinetic properties are similar. However, binding of transition-state analogues is dominated by favorable entropy in VcMTAN and by enthalpy in EcMTAN. Catalytic sites of VcMTAN and EcMTAN in contact with reactants differ by two residues; Ala113 and Val153 in VcMTAN are Pro113 and Ile152, respectively, in EcMTAN. We mutated the VcMTAN catalytic site residues to match those of EcMTAN in anticipation of altering its properties toward EcMTAN. Inhibition of VcMTAN by transition-state analogues required filling both active sites of the homodimer. However, in the Val153Ile mutant or double mutants, transition-state analogue binding at one site caused complete inhibition. Therefore, a single amino acid, Val153, alters the catalytic site cooperativity in VcMTAN. The transition-state analogue affinity and thermodynamics in mutant VcMTAN became even more unlike those of EcMTAN, the opposite of expectations from catalytic site similarity; thus, catalytic site contacts in VcMTAN are unable to recapitulate the properties of EcMTAN. X-ray crystal structures of EcMTAN, VcMTAN, and a multiple-site mutant of VcMTAN most closely resembling EcMTAN in catalytic site contacts show no major protein conformational differences. The overall protein architectures of these closely related proteins are implicated in contributing to the catalytic site differences.

PubMed: 25806409
DOI: 10.1021/bi501487w

Experimental method

X-RAY DIFFRACTION (1.64 Å)