TY - JOUR
T1 - Identification of the five essential histidine residues for peptidylglycine monooxygenase
AU - Yonekura, Hideto
AU - Anzai, Takao
AU - Kato, Ichiro
AU - Furuya, Yasuhito
AU - Shizuta, Satoshi
AU - Takasawa, Shin
AU - Okamoto, Hiroshi
N1 - Funding Information:
We are indebted to Dr. M. Tajima, Dr. T. Iida and Mr. M. Yanagi, Shiseido Basic Research Laboratories, for kindly supplying substrate-affinity gel and, also, for many useful suggestions. This work has been supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture, Japan and Japan Foundation for Applied Enzymology, and is in partial fulfillment by T. A. of the degree of Doctor of Medical Science at Tohoku University. Mr. Brent Bell assisted in preparing the manuscript for publication.
PY - 1996/1/17
Y1 - 1996/1/17
N2 - Peptidylglycine monooxygenase (PGM) is a copper-containing monooxygenase that plays a key role in the peptide C-terminal α-amidation. Comparative analysis of the amino acid sequences of rat, human, bovine and frog PGMs revealed that ten histidines (residues 107, 108, 172, 235, 242, 244, 279, 364, 366 and 367 in rat PGM) are conserved among the four species. We introduced site-directed mutations to the ten histidines of rat PGM and found that the mutation of His-107 → Ala, His-108 → Ala, His-172 → Ala, His-242 → Arg or His-244 → Ala abolished the enzyme activity. The five mutant proteins lacking the enzyme activity bound to a substrate, Phe-Gly-Phe-Gly, as did the wild type PGM. These results along with available evidence indicate that the five histidine residues (His-107, 108, 172, 242 and 244) are essential for PGM activity, acting as copper ligands.
AB - Peptidylglycine monooxygenase (PGM) is a copper-containing monooxygenase that plays a key role in the peptide C-terminal α-amidation. Comparative analysis of the amino acid sequences of rat, human, bovine and frog PGMs revealed that ten histidines (residues 107, 108, 172, 235, 242, 244, 279, 364, 366 and 367 in rat PGM) are conserved among the four species. We introduced site-directed mutations to the ten histidines of rat PGM and found that the mutation of His-107 → Ala, His-108 → Ala, His-172 → Ala, His-242 → Arg or His-244 → Ala abolished the enzyme activity. The five mutant proteins lacking the enzyme activity bound to a substrate, Phe-Gly-Phe-Gly, as did the wild type PGM. These results along with available evidence indicate that the five histidine residues (His-107, 108, 172, 242 and 244) are essential for PGM activity, acting as copper ligands.
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U2 - 10.1006/bbrc.1996.0088
DO - 10.1006/bbrc.1996.0088
M3 - Article
C2 - 8561784
AN - SCOPUS:0030033957
VL - 218
SP - 495
EP - 499
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -