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)

An isoactin analysis was performed on L-[35S]cysteine labeled BC3H1 cells to determine if these smooth muscle-like cells synthesize vascular smooth muscle actin. Three different NH2-terminal peptides were identified on thin layer electrophoretograms of DNase I-purified and trypsin-digested BC3H1 cell actin. Results obtained from secondary digestion with thermolysin or Staphylococcus aureus V8 protease showed that the most acidic NH2-terminal peptide was derived from vascular smooth muscle alpha-isoactin. Treatment of cell monolayers with serum-free medium caused a 3-fold increase in the level of alpha-isoactin expression and a concomitant decrease in the level of non-muscle beta- and gamma-isoactin. Cell-cell contact was required for induction of alpha-isoactin, and the effects of serum depletion on isoactin expression and cell growth were reversible. The intensity of about 11 out of 500 polypeptide spots on two-dimensional gels of BC3H1 cell polypeptides also was influenced by the culture conditions. The finding that smooth muscle isoactin expression was coupled to cell growth conditions indicate the potential usefulness of BC3H1 cells in studies of isoactin expression and utilization during vascular smooth muscle development.
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PMID:Induction of vascular smooth muscle alpha-isoactin expression in BC3H1 cells. 669 10

A fully translated actin biosynthetic intermediate containing N-acetylcysteine at the NH2 terminus has been identified in homogenates of differentiated mouse BC3H1 cerebrovascular smooth muscle cells labeled with L-[35S]cysteine. Thermolysin digestion of the highly acidic NH2-terminal tryptic peptide of this intermediate and electrophoretic analysis of the resulting fragments indicated that the intermediate was a precursor of smooth muscle alpha- isoactin , the major isoactin species in vascular smooth muscle. Carboxypeptidase A digestion of the thermolysin cleavage product corresponding to the first eight amino acid residues of the NH2-terminal tryptic peptide demonstrated an acetylcysteine-glutamate residue at the NH2 terminus. These results imply that the gene for smooth muscle alpha- isoactin , like genes coding for skeletal and cardiac alpha- isoactins , contains a cysteine codon immediately following the initiator methionine codon. Both the methionine and cysteine residues must be removed from the NH2 terminus of the intermediate to yield the mature form of smooth muscle alpha- isoactin . The removal of the cysteine residue in vivo is not direct but apparently involves acetylation of the cysteine and subsequent post-translational cleavage of the resulting acetylcysteine. Such an acetylation-dependent pathway has been demonstrated for removal of cysteine from the NH2 terminus of Drosophila actin synthesized in a cell-free translation system ( Rubenstein , P. A., and Martin, D. J. (1983) J. Biol. Chem. 258, 11354-11360). In vivo pulse-chase experiments indicate that the smooth muscle alpha- isoactin intermediate in BC3H1 cells turns over much more slowly than nonmuscle actin intermediates previously identified in mouse L-cells.
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PMID:A vascular smooth muscle alpha-isoactin biosynthetic intermediate in BC3H1 cells. Identification of acetylcysteine at the NH2 terminus. 672 86

Incubation of purified prostaglandin endoperoxide synthetase from sheep vesicular glands with aspirin results in a covalent binding of the acetyl group of acetylsalicylic acid to the protein. During this acetylation, the cyclooxygenase activity is lost, but not the peroxidase activity. The reaction is completed when almost one acetyl group is bound per polypeptide chain (Mr = 68 000). After proteolysis of [3H]acetyl-protein with pronase, radioactive N-acetylserine was obtained. Originally, however, the hydroxyl group of an internal serine residue in the chain is acetylated. The formation of N-acetylserine can be explained by a rapid O leads to N acetyl shift as soon as the NH2 group of serine is liberated. A radioactive dipeptide was isolated from a thermolysin digest of the [3H]acetyl-enzyme containing phenylalanine and serine, phenylalanine being its N-terminal amino acid. Automatic Edman degradation of native and acetylated enzyme showed that only one polypeptide sequence was present: Ala-Asp-Pro-Gly-Ala-Pro-Ala-Pro-Val-Asn-Pro-X-X-Tyr-. The N-terminal sequence has an apolar character.
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PMID:Acetylation of prostaglandin endoperoxide synthetase with acetylsalicylic acid. 677 69

We present here a new method for inhibiting protein acetylation in a rabbit reticulocyte cell-free protein-synthesizing system. This procedure utilizes S-acetonyl coenzyme A, a nonreactive acetyl-CoA analogue, as an inhibitor of the NH2-terminal protein acetyltransferase in this lysate. With this procedure, we can make, in vitro, Dictyostelium discoideum actin which is 85% nonacetylated but fully translated. With the fully translated but nonacetylated actin as a substrate, the actin can be almost completely acetylated post-translationally in an acetyl-CoA-dependent system after the actin has left the ribosome. Using formylated and nonformylated [35S]Met-tRNAfMet as a source of label and in conjunction with detailed peptide mapping experiments with trypsin and thermolysin, the in vitro acetylation is shown to occur at the NH2 terminus of the newly synthesized actin. Furthermore, the initiator methionine residue, contrary to expectation, is not cleaved off but remains stable for at lest 50 min. thus, in the acetylating reticulocyte lysate system, the primary complete translation product in actin synthesis is Ac-Met-Asp and not Ac-Asp.
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PMID:NH2-terminal acetylation of Dictyostelium discoideum actin in a cell-free protein-synthesizing system. 689 20

A histidine-rich fragment, Cp F5, with a molecular weight of 18,650 was isolated from human ceruloplasmin. It consists of 159 amino acids and contains a possible copper-binding site. The sequence of the first 18 NH2-terminal residues of Cp F5 was determined by automated Edman degradation. Cp F5 was cleaved by cyanogen bromide to produce nine fragments of from 2 to 63 residues. The amino acid sequence of all of the cyanogen bromide fragments was investigated using automated and manual Edman degradation, the fragments being digested with trypsin, chymotrypsin, thermolysin, staphylococcal protease, and pepsin as appropriate. The results, in conjunction with the data on the tryptic peptides reported in the accompanying paper (Kingston, I.B., Kingston, B.L., and Putnam, F.L. (1980) J. Biol. Chem. 255, 2886-2896), establish the complete amino acid sequence of Cp F5.
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PMID:Primary structure of a histidine-rich proteolytic fragment of human ceruloplasmin. I. Amino acid sequence of the cyanogen bromide peptides. 698 29

The complete amino acid sequence of the DNA-binding histone-like protein (Hta) from Thermoplasma acidophilum has been established by sequence studies directly on the protein and on tryptic, chymotryptic, and thermolysin peptides derived from the protein. The sequence of the 89-residue form of HTa is: H2N-Val-Gly-Ile-Ser-Glu-Leu-Ser-Lys-Glu-Val-Ala-Lys-Lys-Ala-Asn-Thr-Thr-Gln-Lys -Val-Ala-Arg-Thr-Val-Ile-Lys-Ser-Phe-Leu-Asp-Glu-Ile-Val-Ser-Glu-Ala-Asn-Gly-Gl y-Gln-Lys-Ile-Asn-Leu-Ala-Gly-Phe-Gly-Ile-Phe-Glu-Arg-Arg-Thr-Gln-Gly-Pro-Arg-L ys-Ala-Arg-Asn-Pro-Gln-Thr-Lys-Lys-Val-Ile-Glu-Val-Pro-Ser-Lys-Lys-Lys-Phe-Val- Phe-Arg-Ala-Ser-Ser-Lys-Ile-Lys-Tyr-Gln-Gln-COOH The molecular weight calculated from the sequence is 9,934. Another form of HTa probably differs only by the presence of an additional residue (methionine) at the NH2 terminus (the calculated molecular weight of this form is 10,065). HTa resembles eukaryotic histones in several ways, including some sequence homology, HTa also shows sequence homology with the Escherichia coli DNA-binding proteins NS1 (or HU-1) and NS2 (or HU-2).
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PMID:A histone-like protein (HTa) from Thermoplasma acidophilum. II. Complete amino acid sequence. 700 26

The peptide alpha 1(III)-CB9 was prepared and purified from human liver, and its amino acid sequence was determined. Automated Edman degradation of the intact peptide and peptides derived from selective cleavage with hydroxylamine and digestions with trypsin, thermolysin, and Staph V8 protease enabled determination of the complete amino acid sequence. The peptide alpha 1(III)-CB9 represents the COOH terminus of the helical (pepsin-resistant) portion of type III collagen and terminates in a Cys-Cys sequence responsible for the intramolecular disulfide cross-linkages with other chains. The present work completes the entire amino acid sequence of the helical (pepsin-resistant) portion of human cirrhotic liver type III collagen consisting of peptides alpha 1-(III)-CB3-7-6-1-8-10-2-4-5-9. The COOH terminus of human liver alpha 1(III) contained two additional triplets which, together with the extra triplet at the NH2 terminus in alpha 1(III)-CB3, make the helical portion of type III collagen longer than alpha 1(I) by nine residues (three Gly-X-Y triplets). The helical region of human liver type III collagen, therefore, consists of 1023 amino acids or 341 triplets.
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PMID:Covalent structure of collagen: amino acid sequence of alpha 1(III)-CB9 from type III collagen of human liver. 701 80

We obtained two fragments of colicin E1 by limited proteolysis with thermolysin. Both of the fragments have an ionophore-like activity. One of the fragments, Th1 (Mr = 18,000), induces an increase in membrane ion permeability comparable with that with whole colicin E1 when assayed with liposomes prepared from soybean phospholipids. This suggests that Th1 is an active fragment of colicin E1. The size of this fragment is about one-third of that of colicin E1 and it was found that the fragment is derived from the COOH-terminal part of colicin E1. A larger fragment, Th2 (Mr = 39,000), was also obtained on thermolysin treatment of colicin E1, which comprises the two-thirds of colicin E1 from the COOH terminus and has a receptor-binding activity in addition to the same ionophore-like activity of Th1. Th2 has the ability to kill osmotic shocked cells but not intact cells, suggesting that the NH2-terminal portion of colicin E1, which was lost in Th2, is responsible for the transport of the colicin molecule through the outer membrane. Thus, the regions which possess or correlate to the ionophore-like, receptor-binding, and transmembrane activities of colicin E1 were assigned to the COOH-terminal, central, and NH2-terminal parts of the molecule, respectively.
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PMID:Assignment of the functional loci in the colicin E1 molecule by characterization of its proteolytic fragments. 704 12

The complete amino acid sequence of the cell attachment domain of human plasma fibronectin (Pierschbacher, M. D., Hayman, E. G., and Ruoslahti, E. (1981) Cell 26, 259-267) has been determined by automated sequential degradation of a peptic fragment comprising this region and of peptides derived from this fragment by digestion with thermolysin, staphylococcal V8 protease, cyanogen bromide cleavage, and partial acid hydrolysis. The fragment contains 108 residues with isoleucine and methionine as the NH2- and carboxyl-terminal amino acids, respectively. No cysteines are present. The calculated molecular weight of the cell attachment fragment, based on the amino acid sequence, is 11,482, which is in good agreement with the molecular weight estimated by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and ultracentrifugation. There are no homologies in this fragment with other published sequences. The implications of the structure of the cell attachment fragment to the molecular mechanism of cell-fibronectin interaction are discussed.
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PMID:The cell attachment domain of fibronectin. Determination of the primary structure. 705 98

The low molecular weight, glutamine-rich storage protein isolated from the seeds of Ricinus communis (castor beans) has been shown to consist of two different polypeptide chains linked by disulfide bond(s). The small subunit is composed of 34 amino acids with a proline at its NH2 terminus, whereas the large subunit contains 61 amino acids with a cyclized glutamine as the NH2-terminal residue. The complete amino acid sequence of both subunits has been determined through characterization of the isolated subunits and selected peptides from trypsin, chymotrypsin, thermolysin, and cyanogen bromide cleavage. The intact protein possesses a large number of glutaminyl and half-cystinyl residues and exhibits sequence heterogeneity as observed from peptide sequences. Comparison of the sequence of this protein and those of other seed proteins indicates some structural similarities between them. The amino acid sequences of the two polypeptide chains of castor bean storage protein are: (formula, see text).
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PMID:Amino acid sequence of small and large subunits of seed storage protein from Ricinus communis. 717 64


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