title: Localization and chemical synthesis of fibronectin peptides with melanoma adhesion and heparin binding activities.
abstract: Tumor cell adhesion to the extracellular matrix is an important consideration in tumor metastasis. Recent results show that multiple adhesion-promoting domains for melanoma cells can be purified from proteolytic digests of fibronectin [McCarthy, J. B., Hagen, S. T., & Furcht, L. T. (1986) J. Cell Biol. 102, 179-188]. Monoclonal antibodies were generated against a tryptic/catheptic 33K heparin binding fragment of fibronectin derived from the carboxyl terminal of the A chain. This region contains a tumor cell adhesion-promoting domain(s). The amino-terminal sequence was determined for this fragment, as well as a tryptic 31K fragment which is located to the carboxyl-terminal side of the 33K heparin binding fragment in A chains of fibronectin. The partial sequence data demonstrate that arginyl-glycyl-aspartyl-serine (RGDS) or the related arginyl-glutamyl-aspartyl-valine (REDV) is not present in the 33K heparin binding fragment, confirming earlier results which demonstrated that cells adhere to this fragment by an RGDS-independent mechanism. Two monoclonal antibodies, termed AHB-1 and AHB-2, recognized epitopes common to heparin binding fragments derived from the carboxyl terminus of both the A and B chains of fibronectin. Monoclonal antibody AHB-2 inhibited melanoma adhesion to the 33K heparin binding fragment of fibronectin in a concentration-dependent manner, whereas monoclonal antibody AHB-1 had no effect on adhesion to this fragment. Neither monoclonal antibody inhibited adhesion to intact fibronectin. However, monoclonal AHB-2 potentiated the inhibitory effect of suboptimal levels of exogenous RGDS on cell adhesion to intact fibronectin. AHB-2 recognized an epitope common to both the A- and B-chain carboxyl-terminal heparin binding region of fibronectin.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455.
authors.forename: J B
authors.initials: JB
authors.lastname: McCarthy
authors.forename: M K
authors.initials: MK
authors.lastname: Chelberg
authors.forename: D J
authors.initials: DJ
authors.lastname: Mickelson
authors.forename: L T
authors.initials: LT
authors.lastname: Furcht
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002460:Cell Line; D005353:Fibronectins / Q000138:chemical synthesis / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000378:metabolism; D011485:Protein Binding; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000378:metabolism*; D014357:Trypsin
chemical_list: D000911:Antibodies, Monoclonal; D005353:Fibronectins; D010446:Peptide Fragments; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic; D006493:Heparin; D014357:Trypsin