Molecular cloning and amino acid sequence of rat kidney aminopeptidase M: a member of a super family of zinc-metallohydrolases.
Using a polyclonal antibody, a partial cDNA clone for rat aminopeptidase M was identified in a lambda gt11 library from rat kidney. A synthetic oligonucleotide probe derived from the sequence of the insert was used to screen a randomly primed lambda gt10 library. This allowed the identification of several overlapping clones encoding the full sequence of the enzyme. The reading frame, 2898 base pairs in length, encodes a 966 amino acid polypeptide. A highly hydrophobic segment, 24 amino acids in length, located close to the aminoterminus, is proposed to serve as the membrane-spanning domain for this membrane-bound enzyme. The sequence includes nine potential N-linked glycosylation sites and one potential sulfation site. In addition, the rat aminopeptidase M sequence contains an eight amino acid consensus sequence believed to serve as the zinc binding domain in a family of zinc-metallohydrolases. Rat aminopeptidase M shows 77% similarity with the recently cloned human enzyme, as well as weaker but significant similarity with aminopeptidase N from E. coli (18%) and with human leukotriene A4 hydrolase (21%).
Malfroy
B
B
Department of Pharmacological Sciences, Genentech, Inc., South San Francisco, CA 94080.
Kado-Fong
H
H
Gros
C
C
Giros
B
B
Schwartz
J C
JC
Hellmiss
R
R
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018826:CD13 Antigens; D003001:Cloning, Molecular; D007668:Kidney / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003012:Microbial Collagenase / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D013820:Thermolysin / Q000235:genetics; D015032:Zinc / Q000378:metabolism*
D000626:Aminopeptidases; D018826:CD13 Antigens; D008666:Metalloendopeptidases; D013820:Thermolysin; D003012:Microbial Collagenase; D015032:Zinc