Sulfatide-binding domain of the laminin A chain

Giulia Taraboletti, C. N. Rao, Henry C. Krutzsch, Lance A. Liotta, David D. Roberts

Research output: Contribution to journalArticle

Abstract

A sulfatide-binding site on the globular end region of the long arm of laminin has been identified. Following proteolytic digestion with thermolysin, an intact fragment of the laminin A chain carboxyl-terminal domain exhibiting sulfatide-binding activity was isolated using gel filtration and heparin affinity chromatography. This fragment is composed of two peptides that are covalently linked by at least one disulfide bond and encompass the carboxyl-terminal 394 amino acids of the A chain. The clusters of charged residues in the primary structure of these fragments are sufficient for heparin-binding activity but not sulfatide binding since reduction and alkylation of the fragments abolished sulfatide binding under conditions in which heparin binding was retained. Thus, sulfatide binding requires an intact three-dimensional structure. The iodinated fragment bound to A2058 melanoma and T47D breast carcinoma cells and could be displaced by the unlabeled fragment. Based on incorporation of [35S] sulfate, both cell lines synthesize sulfated glycolipids that bind to laminin. In agreement with previous data that indicate a synergistic interaction of the sulfatide-binding domain with other laminin-binding sites on melanoma cells during attachment, the isolated sulfatide-binding fragment significantly inhibited interaction of labeled intact laminin with melanoma and breast carcinoma cells in direct binding assays.

Original languageEnglish
Pages (from-to)12253-12258
Number of pages6
JournalJournal of Biological Chemistry
Volume265
Issue number21
Publication statusPublished - Jul 25 1990

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Sulfoglycosphingolipids
Laminin
Heparin
Melanoma
Cells
Binding Sites
Thermolysin
Breast Neoplasms
Affinity chromatography
Alkylation
laminin A
Affinity Chromatography
Disulfides
Sulfates
Gel Chromatography
Digestion
Assays
Thermodynamic properties
Gels
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Taraboletti, G., Rao, C. N., Krutzsch, H. C., Liotta, L. A., & Roberts, D. D. (1990). Sulfatide-binding domain of the laminin A chain. Journal of Biological Chemistry, 265(21), 12253-12258.

Sulfatide-binding domain of the laminin A chain. / Taraboletti, Giulia; Rao, C. N.; Krutzsch, Henry C.; Liotta, Lance A.; Roberts, David D.

In: Journal of Biological Chemistry, Vol. 265, No. 21, 25.07.1990, p. 12253-12258.

Research output: Contribution to journalArticle

Taraboletti, G, Rao, CN, Krutzsch, HC, Liotta, LA & Roberts, DD 1990, 'Sulfatide-binding domain of the laminin A chain', Journal of Biological Chemistry, vol. 265, no. 21, pp. 12253-12258.
Taraboletti G, Rao CN, Krutzsch HC, Liotta LA, Roberts DD. Sulfatide-binding domain of the laminin A chain. Journal of Biological Chemistry. 1990 Jul 25;265(21):12253-12258.
Taraboletti, Giulia ; Rao, C. N. ; Krutzsch, Henry C. ; Liotta, Lance A. ; Roberts, David D. / Sulfatide-binding domain of the laminin A chain. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 21. pp. 12253-12258.
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