Biochemical and functional characterization of aminopeptidase N expressed by human melanoma cells.

Abstract

A cell surface protein expressed on melanoma cells, but not on normal melanocytes, was biochemically and functionally characterized. Microsequencing of the M(r) 143,000 affinity-purified protein revealed amino acid sequence identity to aminopeptidase N (EC 3.4.11.2). In situ expression, indirect immunofluorescence, and Western blotting demonstrated that aminopeptidase N is tightly associated with extracellular matrix components. A specific polyclonal antiserum and the competitive inhibitors of aminopeptidase N, bestatin and amastatin, inhibited invasion of an aminopeptidase N-expressing metastatic melanoma cell line through the reconstituted basement membrane Matrigel in a dose-dependent manner. In vitro digestion of Matrigel with affinity-purified aminopeptidase N revealed an enzyme-sensitive M(r) 160,000 protein. These experiments suggest a role for aminopeptidase N in melanoma invasion of basement membranes.

• Authors: A Menrad, D Speicher, J Wacker, M Herlyn
• Journal: Cancer research (Cancer Res) Vol. 53 Issue 6 Pg. 1450-5 (Mar 15 1993) ISSN: 0008-5472 [Print] United States
• PMID: 8095183 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antibodies, Monoclonal
• Aminopeptidases
• CD13 Antigens

Topics

• Amino Acid Sequence
• Aminopeptidases (chemistry, immunology, metabolism, physiology)
• Antibodies, Monoclonal (immunology)
• CD13 Antigens
• Extracellular Matrix (enzymology)
• Humans
• Melanoma (enzymology, pathology)
• Molecular Weight
• Neoplasm Invasiveness
• Tumor Cells, Cultured