Purification and identification of a novel primitive secretory enzyme catalyzing the hydrolysis of imidazole-related dipeptides in the jawless vertebrate Lethenteron reissneri.
Imidazole-related dipeptides, such as carnosine and anserine, occur widely in skeletal muscles of jawed vertebrates. All of the known enzymes that catalyze the hydrolysis of these dipeptides belong to the M20A metallopeptidase subfamily; two secretory enzymes, serum carnosinase (EC 3.4.13.20) and anserinase (EC 3.4.13.5), and one non-secretory enzyme, cytosolic nonspecific dipeptidase (EC 3.4.13.18). Here we report the enzymatic characterization and molecular identification of an unidentified enzyme, which catalyzes the hydrolysis of these dipeptides, from the skeletal muscle of Far Eastern brook lamprey (Lethenteron reissneri). A 60-kDa subunit protein of the enzyme was purified to near homogeneity. We cloned two M20A genes from the skeletal muscle of Far Eastern brook lamprey; one was a secretory-type gene encoding for the 60-kD protein, and another was a non-secretory-type gene presumably encoding for cytosolic nonspecific dipeptidase. Our findings indicate that the purified enzyme is a N-glycosylated secretory M20A dipeptidase distributed specifically in the jawless vertebrate group, and may be derived from a common ancestor gene between serum carnosinase and anserinase. We propose that this dipeptidase is a novel secretory M20A enzyme and is classified as neither serum carnosinase nor anserinase.
Science of Marine Resources, United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Japan.
Takahiro
T
Oku
Seiichi
S
Ando
Takehiko
T
Hayakawa
Kyoko
K
Baba
Ryuichiro
R
Nishi
Kazuhiro
K
Shiozaki
Shoji
S
Yamada
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055162:Biocatalysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D017931:DNA Primers; D004151:Dipeptides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006868:Hydrolysis; D007093:Imidazoles / Q000378:metabolism*; D007798:Lampreys; D008969:Molecular Sequence Data; D010802:Phylogeny
D017931:DNA Primers; D004151:Dipeptides; D004798:Enzymes; D007093:Imidazoles