Structural features and regulatory properties of the brain glutamate decarboxylases.
It is widely recognized that the two major forms of GAD present in adult vertebrate brains are each composed of two major sequence domains that differ in size and degree of similarity. The amino-terminal domain is smaller and shows little sequence identity between the two forms. This domain is thought to mediate the subcellular targeting of the two GADs. Substantial parts of the amino-terminal domain appear to be exposed and flexible, as shown by proteolysis experiments and the locations of posttranslational modifications. The carboxyl-terminal sequence domain contains the catalytic site and shows substantial sequence similarity between the forms. The interaction of GAD with its cofactor, pyridoxal-5' phosphate (pyridoxal-P), plays a key role in the regulation of GAD activity. Although GAD(65) and GAD(67) interact differently with pyridoxal-P, their cofactor-binding sites contain the same set of nine putative cofactor-binding residues and have the same basic structural fold. Thus the cofactor-binding differences cannot be attributed to fundamental structural differences between the GADs but must result from subtle modifications of the basic cofactor-binding fold. The presence of another conserved motif suggests that the carboxyl-terminal domain is composed of two functional domains: the cofactor-binding domain and a small domain that closes when the substrate binds. Finally, GAD is a dimeric enzyme and conserved features of GADs superfamily of pyridoxal-P proteins indicate the dimer-forming interactions are mediated mainly by the carboxyl-terminal domain.
Martin
D L
DL
Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509, USA. davide.martin@wadsworth.org
Liu
H
H
Martin
S B
SB
Wu
S J
SJ
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002531:Cerebellum / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005968:Glutamate Decarboxylase / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006868:Hydrolysis; D015151:Immunoblotting; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008297:Male; D013058:Mass Spectrometry; D008970:Molecular Weight; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000737:chemistry; D014357:Trypsin
D007527:Isoenzymes; D011994:Recombinant Proteins; D014357:Trypsin; D005968:Glutamate Decarboxylase