Giacin J R, Gilbert S G
Adv Exp Med Biol. 1977;86A:441-71. doi: 10.1007/978-1-4684-3282-4_28.
The effect of structural modification on the enzyme-binding capacity of collagen has been studied using beta-galactosidase (E. coli K12) immobilized to collagen membrane by the impregnation procedure. The apparent steady-state activities of the resultant collagen-enzyme complexes were determined as a means of evaluating the enzyme-binding capacity of the modified collagen. In addition, the amount of enzymic protein bound to the collagen support was determined by the tryptophan content of the complex. The tertiary structure of the collage matrix was modified by cross-linking with the difunctional reagent, glutaraldehyde, and by aging in the dry state. Such structural modifications were found to markedly reduce the enzyme (beta-galactosidase) binding capacity of collagen films. The enzyme-binding capacity of the crosslinked collagen membrane was completely restored by proteolytic enzyme treatment of the aged film but only partly so for the glutaraldehyde treated films. Proteolytic enzymes used to treat a dispersion of collagen microfibrils prior to casting into a membrane also resulted in an increase in enzyme-binding. The effect of structural modification of collagen on enzyme-binding and the locus of enzyme attachment are discussed.
利用通过浸渍法固定在胶原膜上的β-半乳糖苷酶（大肠杆菌K12），研究了结构修饰对胶原酶结合能力的影响。将所得胶原-酶复合物的表观稳态活性作为评估修饰后胶原酶结合能力的一种手段进行测定。此外，通过复合物的色氨酸含量测定结合在胶原载体上的酶蛋白量。通过与双功能试剂戊二醛交联并在干燥状态下老化来修饰胶原基质的三级结构。发现这种结构修饰显著降低了胶原膜的酶（β-半乳糖苷酶）结合能力。通过对老化膜进行蛋白水解酶处理，交联胶原膜的酶结合能力完全恢复，但对于戊二醛处理的膜仅部分恢复。在铸膜前用于处理胶原微纤维分散体的蛋白水解酶也导致酶结合增加。讨论了胶原结构修饰对酶结合的影响以及酶附着的位点。
