cDNA cloning and expression of human glutamyl aminopeptidase (aminopeptidase A).
The murine B-lymphocyte differentiation antigen BP-1/6C3 has been identified as glutamyl aminopeptidase (E-AP), formerly known as aminopeptidase A, the new gene symbol for which is ENPEP. In mice, the enzyme is found on early B-lineage cells and certain stromal cells of the bone marrow and thymus. This ectopeptidase is also expressed by capillary endothelial cells, placenta, and epithelial cells of the intestine and proximal renal tubules. Here we have used a mouse E-AP cDNA to identify the human counterpart in a kidney library. Sequence comparison of the human and mouse cDNAs reveals approximately 80% homology at both nucleotide and predicted amino acid levels. The nucleotide sequence of human E-AP predicts a type II integral membrane protein of 957 amino acids with an 18-amino-acid aminoterminal intracellular domain, and a 22-amino-acid transmembrane domain. The large extracellular carboxyterminal domain contains the zinc-binding motif typical of zinc-dependent metallohydrolases. When the human E-AP cDNA was placed downstream of the SR alpha promoter in an expression vector and transfected into COS-7 cells, the transfected cells exhibited cell surface E-AP activity. A 4.1-kb transcript could be detected in a variety of human tissues, including heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. However, in representative lymphoid leukemias, E-AP transcripts were restricted to pre-B leukemia and were not found in T- and B-cell leukemias. The cDNA cloning and successful expression of human E-AP will allow more precise analysis of its physiological role(s).
Li
L
L
Department of Medicine, University of Alabama at Birmingham 35294.
Wang
J
J
Cooper
M D
MD
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics; D001402:B-Lymphocytes / Q000201:enzymology / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D043384:Glutamyl Aminopeptidase; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000944:Antigens, Differentiation, B-Lymphocyte; D018076:DNA, Complementary; D000626:Aminopeptidases; D043384:Glutamyl Aminopeptidase; D008666:Metalloendopeptidases