Answer
Trypsin is an enzyme that helps us digest protein. In the small intestine, trypsin breaks down proteins, continuing the process of digestion that began in the stomach. It may also be referred to as a proteolytic enzyme, or proteinase. Trypsin is produced by the pancreas in an inactive form called trypsinogen. The trypsinogen enters the small intestine. through the common bile duct and is converted to active trypsin. This active trypsin acts with the other two principal digestive proteinases - pepsin. and chymotrypsin - to break down dietary protein into peptides and amino acids. These amino acids are essential for muscle growth, hormone production and other important bodily functions. Both trypsin and a chymotrypsin are a family of serine proteases, and catalyze the enantioselective hydrolysis of amide and esters. CSPs based on trypsin and a chymotrypsin were introduced by Thelohan et al.4 and Wainer et al.,5 respectively. Trypsin-based CSPs can resolve O, N,O derivatized amino acids that are substrates of the enzyme.4 This means that chiral separations are due to the activity of the enzyme, and that the chiral recognition site could be on the enzyme activity site. a Chymotrypsin-based CSPS can resolve amino acid, amino acid derivatives, dipeptides, and other compounds such as naproxen and aryloxy-propionic acids. chymotrypsin, cleaves peptide bonds selectively. This substrate is an ester rather than an amide, but many proteases will also hydrolyze esters. The products after exposure of eEF-Tu to trypsin for 2 h is a single polypeptide of 43 000 daltons (eEF-Tut) and as yet unidentified polypeptides of Mr less than or equal to 5000. The presence of high glycerol concentrations of GDP in the reaction mixture markedly retards the rate of tryptic cleavage, while GTP has little effect. When eEF-Tu is bound to eucaryotic elongation factor Ts in an eEF-T complex, it is much more resistant to the action of trypsin. The loss of factor activity during tryptic digestion (as measured by its ability to bind aminoacyl-tRNA to 80S (ribosomes) is much slower than the rate of eEF-Tut formation, and 2-h digests containing only eEF-Tut are about 30% as active as the native enzyme. However, no ribosome dependent activity is detectable after purification of eEF-Tut by ion-exchange chromatography, followed by gel filtration. Purified eEF-Tut binds guanine nucleotides, although with diminished activity compared with that of eEF-Tu. Amino-terminal sequence analyses of eEF-Tut reveal a striking sequence homology with the functionally related factor from Escherichia coli (EF-Tu). The first four residues of eEF Tut, Gly-lle-Thr-lle, are identical with the first four residues of a 37 000-dalton tryptic fragment of E. coli EF-Tu, and other homologies are evident in the first twelve amino-terminal residues of the corresponding tryptic fragments. C3c and C3d fragments were prepared in pure form from trypsin-digested human C3, and the individual chains of tryptic C3c were isolated by gel filtration on Sepharose 4B in 6 M guanidinium hydrochloride. No low mol. wt (Mr) fragments were identified. The polypeptide chains were characterized with regard to Mr, amino acid composition and N-terminal amino acid sequence. Tryptic C3c consisted of one fragment from the B-chain (Mr 64,000) and two from the a' chain (Mr 40,000 and 23,000). The B-chain fragment was derived from the C-terminal part of the chain, and the 23,000-Mr component constituted the amino terminal end of the a-chain. The 40,000-Mtr fragment emanated from the C-terminal end of the a-chain. Tryptic C3d displayed microheterogeneity on polyacrylamide gel electrophoresis in sodium dodecyl sulfate, but possessed a homogeneous N-terminal, identical to that described by Tack etal. (1980) (proc. natn. Acad. Sci. U.S.A.77, 5764-5768). By utilization of antisera against subunits of C3 and C3c in immunoblotting a degradation scheme for C3 by trypsin was proposed and the positions of the fragments in the intact molecule indicated.
Work Step by Step
Trypsin is an enzyme that helps us digest protein. In the small intestine, trypsin breaks down proteins, continuing the process of digestion that began in the stomach. It may also be referred to as a proteolytic enzyme, or proteinase. Trypsin is produced by the pancreas in an inactive form called trypsinogen. The trypsinogen enters the small intestine. through the common bile duct and is converted to active trypsin. This active trypsin acts with the other two principal digestive proteinases - pepsin. and chymotrypsin - to break down dietary protein into peptides and amino acids. These amino acids are essential for muscle growth, hormone production and other important bodily functions. Both trypsin and a chymotrypsin are a family of serine proteases, and catalyze the enantioselective hydrolysis of amide and esters. CSPs based on trypsin and a chymotrypsin were introduced by Thelohan et al.4 and Wainer et al.,5 respectively. Trypsin-based CSPs can resolve O, N,O derivatized amino acids that are substrates of the enzyme.4 This means that chiral separations are due to the activity of the enzyme, and that the chiral recognition site could be on the enzyme activity site. a Chymotrypsin-based CSPS can resolve amino acid, amino acid derivatives, dipeptides, and other compounds such as naproxen and aryloxy-propionic acids. chymotrypsin, cleaves peptide bonds selectively. This substrate is an ester rather than an amide, but many proteases will also hydrolyze esters. The products after exposure of eEF-Tu to trypsin for 2 h is a single polypeptide of 43 000 daltons (eEF-Tut) and as yet unidentified polypeptides of Mr less than or equal to 5000. The presence of high glycerol concentrations of GDP in the reaction mixture markedly retards the rate of tryptic cleavage, while GTP has little effect. When eEF-Tu is bound to eucaryotic elongation factor Ts in an eEF-T complex, it is much more resistant to the action of trypsin. The loss of factor activity during tryptic digestion (as measured by its ability to bind aminoacyl-tRNA to 80S (ribosomes) is much slower than the rate of eEF-Tut formation, and 2-h digests containing only eEF-Tut are about 30% as active as the native enzyme. However, no ribosome dependent activity is detectable after purification of eEF-Tut by ion-exchange chromatography, followed by gel filtration. Purified eEF-Tut binds guanine nucleotides, although with diminished activity compared with that of eEF-Tu. Amino-terminal sequence analyses of eEF-Tut reveal a striking sequence homology with the functionally related factor from Escherichia coli (EF-Tu). The first four residues of eEF Tut, Gly-lle-Thr-lle, are identical with the first four residues of a 37 000-dalton tryptic fragment of E. coli EF-Tu, and other homologies are evident in the first twelve amino-terminal residues of the corresponding tryptic fragments. C3c and C3d fragments were prepared in pure form from trypsin-digested human C3, and the individual chains of tryptic C3c were isolated by gel filtration on Sepharose 4B in 6 M guanidinium hydrochloride. No low mol. wt (Mr) fragments were identified. The polypeptide chains were characterized with regard to Mr, amino acid composition and N-terminal amino acid sequence. Tryptic C3c consisted of one fragment from the B-chain (Mr 64,000) and two from the a' chain (Mr 40,000 and 23,000). The B-chain fragment was derived from the C-terminal part of the chain, and the 23,000-Mr component constituted the amino terminal end of the a-chain. The 40,000-Mtr fragment emanated from the C-terminal end of the a-chain. Tryptic C3d displayed microheterogeneity on polyacrylamide gel electrophoresis in sodium dodecyl sulfate, but possessed a homogeneous N-terminal, identical to that described by Tack etal. (1980) (proc. natn. Acad. Sci. U.S.A.77, 5764-5768). By utilization of antisera against subunits of C3 and C3c in immunoblotting a degradation scheme for C3 by trypsin was proposed and the positions of the fragments in the intact molecule indicated.
