Pre-steady-state kinetic and structural analysis of interaction of methionine γ-lyase from Citrobacter freundii with inhibitors.
Methionine γ-lyase (MGL) catalyzes the γ-elimination of l-methionine and its derivatives as well as the β-elimination of l-cysteine and its analogs. These reactions yield α-keto acids and thiols. The mechanism of chemical conversion of amino acids includes numerous reaction intermediates. The detailed analysis of MGL interaction with glycine, l-alanine, l-norvaline, and l-cycloserine was performed by pre-steady-state stopped-flow kinetics. The structure of side chains of the amino acids is important both for their binding with enzyme and for the stability of the external aldimine and ketimine intermediates. X-ray structure of the MGL·l-cycloserine complex has been solved at 1.6 Å resolution. The structure models the ketimine intermediate of physiological reaction. The results elucidate the mechanisms of the intermediate interconversion at the stages of external aldimine and ketimine formation.
From the Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, the Department of Natural Sciences, Novosibirsk State University, Novosibirsk 630090.
Nikita A
NA
Kuznetsov
Nesmeyanov Institute of Organoelement Compounds and.
Nicolai G
NG
Faleev
From the Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, the Department of Natural Sciences, Novosibirsk State University, Novosibirsk 630090.
Alexandra A
AA
Kuznetsova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, and.
Elena A
EA
Morozova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, and.
Svetlana V
SV
Revtovich
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, and.
Natalya V
NV
Anufrieva
the Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
Alexei D
AD
Nikulin
From the Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, the Department of Natural Sciences, Novosibirsk State University, Novosibirsk 630090, fedorova@niboch.nsc.ru.
Olga S
OS
Fedorova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, and tvd@eimb.ru.
Tatyana V
TV
Demidkina
D000409:Alanine / Q000737:chemistry; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D013437:Carbon-Sulfur Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D020134:Catalytic Domain; D016971:Citrobacter freundii / Q000737:chemistry* / Q000201:enzymology; D018360:Crystallography, X-Ray; D003523:Cycloserine / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005998:Glycine / Q000737:chemistry; D007097:Imines / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D011732:Pyridoxal Phosphate / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013816:Thermodynamics; D014633:Valine / Q000031:analogs & derivatives / Q000737:chemistry
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D007097:Imines; D011994:Recombinant Proteins; D011732:Pyridoxal Phosphate; D003523:Cycloserine; C005313:norvaline; D013437:Carbon-Sulfur Lyases; C021558:L-methionine gamma-lyase; D014633:Valine; D003545:Cysteine; D000409:Alanine; D005998:Glycine
Enzyme Inhibitor; Enzyme Structure; Methionine γ-Lyase; Pre-steady-state Kinetics; Pyridoxal Phosphate; Structure of Pyridoxal 5′-Phosphate-Cycloserine Derivative; Substrate Specificity