Enzymatic Reaction:
Trypsin is a pancreatic serine protease with substrate specificity based upon positively charged lysine and arginine side chains (Brown and Wold 1973). The enzyme in excreted by the pancreas and takes part in the digestion of food proteins and other biological processes. Trypsin is a medium-sized globular protein and is produced as an inactive proenzyme, trypsinogen (Chen et al. 2009).
Specificity:
Trypsin cleaves peptides on the C-terminal side of lysine and arginine amino acid residues. If a proline residue is on the carboxyl side of the cleavage site, the cleavage will not occur. If an acidic residue is on either side of the cleavage site, the rate of hydrolysis has been shown to be slower.
Applications:
Tissue dissociation, especially when combined with other enzymes such as collagenase, and elastase
Cell harvesting by “trypsinization”
Mitochondria isolation
in vitro studies of proteins
Removing monolayers of cells from plastic and glass
Various hemagglutination procedures
Sample preparation for flow cytometric DNA analysis
Tryptic mapping
Fingerprinting and sequencing work
Environmental monitoring
Reduction of cell density in tissue culture
Subculturing cells
Cleavage fusion proteins
Generating glycopeptides from purified glycoproteins
胰蛋白酶切割赖氨酸和精氨酸氨基酸残基 C 端侧的肽。如果脯氨酸残基位于切割位点的羧基侧,则不会发生切割。如果酸性残基位于裂解位点的任一侧,则水解速率已显示较慢。
胰蛋白酶相关研究:
细胞分离优化系统
胰凝乳蛋白酶
胶原酶
脱氧核糖核酸酶 I
内切蛋白酶Lys-C
肝细胞分离系统
透明质酸酶
新生儿心肌细胞分离系统
中性蛋白酶(分散酶)
木瓜蛋白酶解离系统
蛋白酶K
胰蛋白酶抑制剂
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