van den Bosch Edith, Gielens Constant
Laboratory for Biochemistry, Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200 G, B-3001 Leuven, Belgium.
Int J Biol Macromol. 2003 Sep;32(3-5):129-38. doi: 10.1016/s0141-8130(03)00046-1.
Four gelatin types (A, B, C and AB), two different samples of each, were subjected to temperature treatments with the incubation temperature, incubation time, gelatin concentration and solvent (type and concentration of salt ions and pH) as variables. Degradation was studied by means of fast protein liquid chromatography and sodium dodecylsulphate polyacrylamide gel electrophoresis. All the variables tested seemed to be critical. Addition of a protease inhibitor cocktail confirmed that the observed degradation was not due to the action of proteases.Fluorescence measurements indicated that during the temperature treatment pentosidine and pyridinoline cross-links can be broken, while the cleavage of peptide bonds was verified by ninhydrin tests and N-terminal amino-acid analyses with phenyl isothiocyanate.
四种明胶类型（A、B、C和AB），每种类型有两个不同的样品，以孵育温度、孵育时间、明胶浓度和溶剂（盐离子类型和浓度以及pH值）作为变量进行温度处理。通过快速蛋白质液相色谱法和十二烷基硫酸钠聚丙烯酰胺凝胶电泳研究降解情况。所有测试变量似乎都很关键。添加蛋白酶抑制剂混合物证实观察到的降解不是由蛋白酶的作用引起的。荧光测量表明，在温度处理过程中，戊糖苷和吡啶啉交联键可以断裂，而肽键的断裂通过茚三酮试验和用异硫氰酸苯酯进行的N端氨基酸分析得到证实。
