ADP-ribosylation of integrin alpha7 modulates the binding of integrin alpha7beta1 to laminin.
The extracellular domain of integrin alpha7 is ADP-ribosylated by an arginine-specific ecto-ADP-ribosyltransferase after adding exogenous NAD+ to intact C2C12 skeletal muscle cells. The effect of ADP-ribosylation on the structure or function of integrin alpha7beta1 has not been explored. In the present study, we show that ADP-ribosylation of integrin alpha7 takes place exclusively in differentiated myotubes and that this post-translational modification modulates the affinity of alpha7beta1 dimer for its ligand, laminin. ADP-ribosylation in the 37-kDa 'stalk' region of alpha7 that takes place at micromolar NAD+ concentrations increases the binding of the alpha7beta1 dimer to laminin. Increased in vitro binding of integrin alpha7beta1 to laminin after ADP-ribosylation of the 37-kDa fragment of alpha7 requires the presence of Mn2+ and it is not observed in the presence of Mg2+. In contrast, ADP-ribosylation of the 63-kDa N-terminal region comprising the ligand-binding site of alpha7 that occurs at approx. 100 microM NAD+ inhibits the binding of integrin alpha7beta1 to laminin. Furthermore, incubation of C2C12 myotubes with NAD+ increases the expression of an epitope on integrin beta1 subunit recognized by monoclonal antibody 9EG7. We discuss our results based on the current models of integrin activation. We also hypothesize that ADP-ribosylation may represent a mechanism of regulation of integrin alpha7beta1 function in myofibres in vivo when the continuity of the membrane is compromised and NAD+ is available as a substrate for ecto-ADP-ribosylation.
Zhao
Zhefeng
Z
Department of Biochemistry, Kansas State University, 104 Willard Hall, Manhattan, KS 66506, USA.
Gruszczynska-Biegala
Joanna
J
Zolkiewska
Anna
A
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism*; D002413:Cations, Divalent / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D039001:Integrin alpha Chains / Q000737:chemistry / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D051379:Mice; D009243:NAD / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects
D015703:Antigens, CD; D002413:Cations, Divalent; D039001:Integrin alpha Chains; D016023:Integrins; D007797:Laminin; C085725:integrin alpha7; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D008345:Manganese; D008274:Magnesium; C082908:integrin alpha7beta1