Chimeras of the integrin beta subunit mid-region reveal regions required for heterodimer formation and for activation.
A central region of the beta2 integrin subunit, RN (residues D300 to C459), was replaced by the equivalent sequences from beta1 and beta7 to give the chimeras beta2RN1 and beta2RN7. Whilst the former construct failed to form heterodimer at the cell surface with alphaL, the later of these could be expressed together with the alphaL subunit to form a variant LFA-1. Based on recent modelling work, the RN region consists of two parts, one is the C-terminal end of the putative A-domain (RB, residues D300 to A359), and the other the mid-region (BN, residues Y360 to C459). Chimeras exchanging the two component regions were made. Of the four resultant chimeras, only the beta2RB1 chimera failed to support LFA-1 expression. Thus the beta1 specific residues of this region affect the interaction with the alphaL subunit. Whereas the alphaL/beta2RB7 LFA-1 variant is wildtype like with respect to ICAM-1 adhesion, the alphaLbeta2BN1 and alphaLbeta2BN7, as well as the alphaLbeta2RN7, variants are more adhesive than the wildtype. These results suggest that an authentic beta2 mid-region is, in part, required for maintaining the LFA-1 in a resting state.
Hyland
R H
RH
The MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, UK.
Douglass
W A
WA
Tan
S M
SM
Law
S K
SK
D000595:Amino Acid Sequence; D000818:Animals; D018821:CD18 Antigens / Q000737:chemistry* / Q000378:metabolism; D019556:COS Cells; D002448:Cell Adhesion; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D018821:CD18 Antigens; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D018799:Intercellular Adhesion Molecule-1