The partial amino acid sequence of the NAD(+)-dependent glutamate dehydrogenase of Clostridium symbiosum: implications for the evolution and structural basis of coenzyme specificity.
The amino acid sequence is reported for CNBr and tryptic peptide fragments of the NAD(+)-dependent glutamate dehydrogenase of Clostridium symbiosum. Together with the N-terminal sequence, these make up about 75% of the total sequence. The sequence shows extensive similarity with that of the NADP(+)-dependent glutamate dehydrogenase of Escherichia coli (52% identical residues out of the 332 compared) allowing confident placing of the peptide fragments within the overall sequence. This demonstrated sequence similarity with the E. coli enzyme, despite different coenzyme specificity, is much greater than the similarity (31% identities) between the GDH's of C. symbiosum and Peptostreptococcus asaccharolyticus, both NAD(+)-linked. The evolutionary implications are discussed. In the 'fingerprint' region of the nucleotide binding fold the sequence Gly X Gly X X Ala is found, rather than Gly X Gly X X Gly. The sequence found here has previously been associated with NADP+ specificity and its finding in a strictly NAD(+)-dependent enzyme requires closer examination of the function of this structural motif.
Lilley
K S
KS
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, UK.
Baker
P J
PJ
Britton
K L
KL
Stillman
T J
TJ
Brown
P E
PE
Moir
A J
AJ
Engel
P C
PC
Rice
D W
DW
Bell
J E
JE
Bell
E
E
D000595:Amino Acid Sequence; D005075:Biological Evolution; D003013:Clostridium / Q000201:enzymology*; D003488:Cyanogen Bromide; D005969:Glutamate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009243:NAD / Q000502:physiology; D012689:Sequence Homology, Nucleic Acid; D014357:Trypsin
D009243:NAD; D005969:Glutamate Dehydrogenase; D014357:Trypsin; D003488:Cyanogen Bromide