This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hibino, S. Articles by Kleinman, H. K. Search for Related Content PubMed PubMed Citation Articles by Hibino, S. Articles by Kleinman, H. K.

Identification of an Active Site on the Laminin 5 Chain Globular Domain That Binds to CD44 and Inhibits Malignancy

¹ Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, NIH, Bethesda, Maryland; ² Fourth Department of Internal Medicine, Nippon Medical School, Tokyo, Japan; ³ Respiratory Division of Internal Medicine, Tokyo Metropolitan Komagome Hospital, Tokyo, Japan; and ⁴ Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan

The laminin 5 chain is a component of laminin-10 (5ß11) and -11 (5ß21). In this study, we have screened 113 overlapping synthetic peptides from the laminin 5 globular domain (G-domain) for cell attachment activity with B16-F10 cells using peptide-coated dishes. Eleven attachment-active peptides were identified. In vivo experimental B16-F10 pulmonary metastasis and primary tumor growth assays found that 4 of the 11 peptides inhibited tumor metastasis and growth and increased apoptosis. These four peptides also blocked tumor cell migration, invasion, and angiogenesis. Two of the peptides were highly homologous and showed significant similarity to sequences in collagens. We sought to identify the B16-F10 cell surface receptors for each of the four active peptides using peptide affinity chromatography. Only one peptide recognized a cell surface protein. Peptide A5G27 (RLVSYNGIIFFLK, residues 2892–2904) bound a diffuse M_r 120,000–180,000 band that eluted with 2 M NaCl. Glycosidase digestion of the 2 M eluate yielded protein bands of M_r 90,000 and 60,000 that reacted in Western blot analysis with antibodies to CD44. Immunoprecipitation of the A5G27-bound membrane proteins with various cell surface proteoglycan antibodies confirmed CD44 as the surface receptor for A5G27. Finally, attachment assays to A5G27 in the presence of soluble glycosaminoglycans (GAGs) identified the GAGs of CD44 as the binding sites for A5G27. Our results suggest that A5G27 binds to the CD44 receptor of B16-F10 melanoma cells via the GAGs on CD44 and, thus, inhibits tumor cell migration, invasion, and angiogenesis in a dominant-negative manner.

This article has been cited by other articles:

				H. L. Goel, L. Moro, J. E. Murphy-Ullrich, C.-C. Hsieh, C.-L. Wu, Z. Jiang, and L. R. Languino {beta}1 Integrin Cytoplasmic Variants Differentially Regulate Expression of the Antiangiogenic Extracellular Matrix Protein Thrombospondin 1 Cancer Res., July 1, 2009; 69(13): 5374 - 5382. [Abstract] [Full Text] [PDF]

				A. Akalu, J. M. Roth, M. Caunt, D. Policarpio, L. Liebes, and P. C. Brooks Inhibition of Angiogenesis and Tumor Metastasis by Targeting a Matrix Immobilized Cryptic Extracellular Matrix Epitope in Laminin Cancer Res., May 1, 2007; 67(9): 4353 - 4363. [Abstract] [Full Text] [PDF]

				T. C. Sroka, M. E. Pennington, and A. E. Cress Synthetic D-amino acid peptide inhibits tumor cell motility on laminin-5 Carcinogenesis, September 1, 2006; 27(9): 1748 - 1757. [Abstract] [Full Text] [PDF]

				S. Hibino, M. Shibuya, M. P. Hoffman, J. A. Engbring, R. Hossain, M. Mochizuki, S. Kudoh, M. Nomizu, and H. K. Kleinman Laminin {alpha}5 Chain Metastasis- and Angiogenesis-Inhibiting Peptide Blocks Fibroblast Growth Factor 2 Activity by Binding to the Heparan Sulfate Chains of CD44 Cancer Res., November 15, 2005; 65(22): 10494 - 10501. [Abstract] [Full Text] [PDF]