Owens R J, Baralle F E
EMBO J. 1986 Nov;5(11):2825-30. doi: 10.1002/j.1460-2075.1986.tb04575.x.
The collagen-binding domain of human fibronectin has been expressed as a cro/beta-galactosidase fusion protein in Escherichia coli. The hybrid polypeptide was recognized by an anti-(human plasma fibronectin) serum and bound specifically to gelatin-Sepharose. The collagen-binding region was subdivided by constructing a series of overlapping bacterial expression plasmids. The fusion proteins produced by these constructs were analysed for gelatin-binding activity. The results indicate that the binding site lies within an approximately 12.5 kd fragment of fibronectin, and show that the following 14 amino acid sequence is critical for gelatin-binding activity: Ala-Ala-His-Glu-Glu-Ile-Cys-Thr-Thr-Asn-Glu-Gly-Val-Met. This sequence links the second type II homology unit with the adjacent type I repeat in the amino-terminal third of the fibronectin molecule.
人纤连蛋白的胶原结合结构域已在大肠杆菌中作为一种cro/β-半乳糖苷酶融合蛋白表达。该杂合多肽可被抗(人血浆纤连蛋白)血清识别,并能特异性结合明胶-琼脂糖。通过构建一系列重叠的细菌表达质粒对胶原结合区域进行了细分。分析了这些构建体产生的融合蛋白的明胶结合活性。结果表明,结合位点位于纤连蛋白约12.5kd的片段内,并且表明以下14个氨基酸序列对明胶结合活性至关重要:Ala-Ala-His-Glu-Glu-Ile-Cys-Thr-Thr-Asn-Glu-Gly-Val-Met。该序列在纤连蛋白分子氨基末端三分之一处将第二个II型同源单位与相邻的I型重复序列相连。