Defining fibronectin's cell adhesion synergy site by site-directed mutagenesis.

被引:154
作者
Redick, SD [1 ]
Settles, DL [1 ]
Briscoe, G [1 ]
Erickson, HP [1 ]
机构
[1] Duke Univ, Med Ctr, Dept Cell Biol, Durham, NC 27710 USA
关键词
RGD; integrin; fibronectin type III; alpha; 5; beta; 1;
D O I
10.1083/jcb.149.2.521
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Fibronectin's RGD-mediated binding to the alpha 5 beta 1 integrin is dramatically enhanced by a synergy site within fibronectin III domain 9 (FN9). Guided by the crystal structure of the cell-binding domain, we selected amino acids in FN9 that project in the same direction as the RGD, presumably toward the integrin, and mutated them to alanine. R1379 in the peptide PHSRN, and the nearby R1374 have been shown previously to be important for alpha 5 beta 1-mediated adhesion (Aota, S., M. Nomizu, and K.M. Yamada. 1994, J. Biol. Chem, 269:24756-24761), Our more extensive set of mutants showed that R1379 is the key residue in the synergistic effect, but other residues contribute substantially, R1374A decreased adhesion slightly by itself, but the double mutant R1374A-R1379A was significantly less adhesive than R1379A alone. Single mutations of R1369A, R1371A, T1385A, and N1386A had negligible effects on cell adhesion, but combining these substitutions either with R1379A or each other gave a more dramatic reduction of cell adhesion. The triple mutant R1374A/P1376A/R1379A had no detectable adhesion activity. We conclude that, in addition to the R of the PHRSN peptide, other residues on the same face of FN9 are required for the full synergistic effect. The integrin-binding synergy site is a much more extensive surface than the small linear peptide sequence.
引用
收藏
页码:521 / 527
页数:7
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