Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Utilization of a soluble integrin-alkaline phosphatase chimera to characterize integrin alpha 8 beta 1 receptor interactions with tenascin: Murine alpha 8 beta 1 binds to the rgd site in tenascin-c fragments, but not to native tenascin-c

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Denda, S.
  • Mueller, Ulrich
  • Crossin, Kathryn
  • Erickson, H. P.
  • Reichardt, L. F.

publication date

  • 1998

journal

  • Biochemistry  Journal

abstract

  • The integrin alpha 8 beta 1 has been reported to bind to fibronectin, vitronectin, and tenascin-C in cell adhesion or neurite outgrowth assays. Here, we describe cDNA cloning of the murine alpha 8 subunit, purification of a recombinant soluble heterodimer consisting of the extracellular domains of the murine alpha 8 and beta1 subunits, and development of a sensitive binding assay using a modified form of this heterodimer fused to alkaline phosphatase (AP). In binding assays, the purified alpha 8 beta 1-AP chimera exhibited the same divalent ion requirements for activation and binding specificity as cell surface alpha 8 beta 1: in the presence of Mn2+ it bound to fibronectin and vitronectin in an RGDS-peptide inhibitable manner. Contrary to previous reports, we found no evidence that alpha 8 beta 1, expressed on K562 cells or as an AP chimera, interacts strongly with native tenascin-C. In binding, adhesion, and spreading assays, significant interactions were observed only to short fragments of tenascin-C containing the third fibronectin type III repeat which contains an RGD sequence. Full length tenascin-C and longer fragments containing this repeat did not appear to serve as ligands, implying that the RGD site in native tenascin-C is a cryptic binding site for this integrin, exposed by removal of adjacent domains. Soluble integrin-AP chimeras should be generally useful for identifying and characterizing integrin interactions with ligands.

subject areas

  • Alkaline Phosphatase
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Adhesion
  • Chickens
  • Cloning, Molecular
  • Dimerization
  • Humans
  • Integrin alpha Chains
  • Integrins
  • Mice
  • Molecular Sequence Data
  • Oligopeptides
  • Peptide Fragments
  • Protein Binding
  • Recombinant Fusion Proteins
  • Repetitive Sequences, Nucleic Acid
  • Solubility
  • Tenascin
scroll to property group menus

Identity

PubMed Central ID

  • PMC2710129

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi9727489

PubMed ID

  • 9548928
scroll to property group menus

Additional Document Info

start page

  • 5464

end page

  • 5474

volume

  • 37

issue

  • 16

©2019 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support