Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.27 (thermolysin)
 1,894 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The complete covalent structure of a small, basic protein with cardiotoxic activity is described. This has been isolated from the venom of Naja nigricollis by gel filtration on Sephadex G-75 and gradient ion exchange chromatography on Bio-Rex 70. The cardiotoxin, molecular weight 6806 from amino acid composition, consists of 60 amino acids, cross-linked by four disulfide bridges, connecting 3-21, 14-38, 42-53, and 54-59. The protein contains one residue of tryptophan, phenylalanine, and glutamic acid, two residues of arginine and tyrosine, four residues of methionine, and nine residues of lysine. Histidine is absent. The chymotryptic peptides of the oxidized and S-carboxymethylated protein were isolated by gel filtration on Sephadex G-25 and zone electrophoresis on a cellulose column. The sequence was determined by Edman degradation, using the (manual) direct phenylthiohydantoin method and with the use of carboxypeptidase A. Disulfide pairing was determined on thermolysin cleaved peptides from the native protein. The sequence is shown to be homologous to other cardiotoxins and a lytic factor from snake venoms and also shows homology, both in sequence and disulfide pairing to neurotoxins. A partial reduction experiment in the absence of denaturing agent using 14-C-labeled iodoacetic acid as S-carboxymethylating agent shows that disulfide bonds 14-38 and 42-53 were reduced fastest followed marginally by 54-59, and then bond 3-21.
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PMID:The complete covalent structure of a cardiotoxin from the venom of Naja nigricollis (African black-necked spitting cobra). 114 81

The assignment of five disulfide bonds in the alpha subunit of human chorionic gonadotropin (hCG) using partial reduction and S-[14C]carboxymethylation has been reported earlier (Mise, T., and Bahl, O. P. (1980) J. Biol. Chem. 255, 8516-8522). Employing a similar approach, we have determined the locations of six disulfide bonds in hCG-beta. Two partially reduced and S-[14C]carboxymethylated hCG-beta derivatives, DS1.4-hCG-beta and DS3.4-hCG-beta in which on the average 1.4 and 3.4 disulfide bonds were modified, respectively, were prepared. The 14C-labeled derivatives were then completely reduced and S-carboxymethylated with nonradioactive iodoacetic acid and subjected to hydrolysis with trypsin. The radioactive peptides were purified by gel filtration and high voltage paper electrophoresis. The tryptic peptides containing two or more S-[14C]carboxymethylcysteines were further degraded using various proteolytic enzymes such as thermolysin, carboxypeptidase A and Y, cathespin C, and subtilisin to obtain individual S-[14C]carboxymethylcysteine-containing peptides. From the specific radioactivities of S-[14C]carboxymethylcysteines in DS3.4-hCG-beta, four out of six disulfide bonds, 9-90, 26-110, 34-88, and 93-100 were assigned. Similar data from DS1.4-hCG-beta gave the locations of the other two disulfide bonds, 23-72 and 38-57, while confirming the locations of four disulfide bonds derived from the radioactivity distribution in DS3.4-hCG-beta. Thus, all six disulfide bonds in hCG-beta have been located. The results of controlled reduction and S-[14C]alkylation also indicate that disulfide bond 93-100 is the most reactive, followed by disulfide bond 26-110, and that the least reactive among all is 34-88.
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PMID:Assignment of disulfide bonds in the beta subunit of human chorionic gonadotropin. 724 Feb 31

Ovalbumin isolated from eggs of the Japanese quail, C. c. japonica, was subjected to limited proteolysis by subtilisin to give plakalbumin and then fractionated on Sephadex G75 in acid-urea to give plakalbumin S-protein and S-peptide. The plakalbumin peptide was recovered, oxidized with performic acid, and the sequence of amino acids determined from the peptides formed by enzyme digestion. There were two cysteine residues in the 33-residue sequence. The ovalbumin was also oxidized with performic acid and digested with thermolysin and pepsin before isolating, from a sulfonated polystyrene column, the acidic cysteic acid peptides, as well as acetylated N-terminal peptides and phosphorylated peptides, and determining their amino acid sequence. Additional peptide sequences containing cysteine or half-cystine were characterized. Quail ovalbumin was reduced and carboxymethylated with [2-14C]iodoacetic acid. Peptides containing labelled S-carboxymethylcysteine residues were isolated from thermolytic digests of the carboxymethylated ovalbumin by paper ionophoresis and chromatography. Their amino acid sequence was determined and five different sequences involving labelled S-carboxymethylcysteine residues were established. The presence of two half-cystine residues and the location of the disulfide bond were shown by blocking the cysteine residues with non-radioactive iodoacetic acid, reducing the disulfide bond and labelling the half-cystine residues with [2-14C]iodoacetic acid. After thermolytic digestion of the protein, radioactive peptides were isolated by paper ionophoresis and chromatography. These studies have thus shown that quail ovalbumin contains one cystine residue and three cysteine residues, which is one residue of cysteine less than in ovalbumin from the hen (Gallus gallus domesticus). There is strong homology in the amino acid sequences of hen ovalbumin and quail ovalbumin determined in these investigations.
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PMID:Amino acid sequences containing cysteine or cystine residues in ovalbumin from eggs of the quail Coturnix coturnix japonica. 734 Jul 65