Evidence that distinct states of the integrin alpha6beta1 interact with laminin and an ADAM.
Integrins can exist in different functional states with low or high binding capacity for particular ligands. We previously provided evidence that the integrin alpha6beta1, on mouse eggs and on alpha6-transfected cells, interacted with the disintegrin domain of the sperm surface protein ADAM 2 (fertilin beta). In the present study we tested the hypothesis that different states of alpha6beta1 interact with fertilin and laminin, an extracellular matrix ligand for alpha6beta1. Using alpha6-transfected cells we found that treatments (e.g., with phorbol myristate acetate or MnCl2) that increased adhesion to laminin inhibited sperm binding. Conversely, treatments that inhibited laminin adhesion increased sperm binding. Next, we compared the ability of fluorescent beads coated with either fertilin beta or with the laminin E8 fragment to bind to eggs. In Ca2+-containing media, fertilin beta beads bound to eggs via an interaction mediated by the disintegrin loop of fertilin beta and by the alpha6 integrin subunit. In Ca2+-containing media, laminin E8 beads did not bind to eggs. Treatment of eggs with phorbol myristate acetate or with the actin disrupting agent, latrunculin A, inhibited fertilin bead binding, but did not induce laminin E8 bead binding. Treatment of eggs with Mn2+ dramatically increased laminin E8 bead binding, and inhibited fertilin bead binding. Our results provide the first evidence that different states of an integrin (alpha6beta1) can interact with an extracellular matrix ligand (laminin) or a membrane-anchored cell surface ligand (ADAM 2).
Chen
M S
MS
Department of Cell Biology, University of Virginia, Charlottesville, Virginia 22908, USA.
Almeida
E A
EA
Huovila
A P
AP
Takahashi
Y
Y
Shaw
L M
LM
Mercurio
A M
AM
White
J M
JM
D051722:ADAM Proteins; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019483:Disintegrins / Q000235:genetics / Q000378:metabolism; D005260:Female; D000072196:Fertilins; D066298:In Vitro Techniques; D039121:Integrin alpha6beta1; D016023:Integrins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism; D014162:Transfection; D015053:Zygote / Q000254:growth & development