Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The complete amino acid sequence of mitochondrial glutamic oxaloacetic transaminase from rat liver was determined. Sequence analyses were performed on the 12 cyanogen bromide peptides isolated after cleavage with cyanogen bromide. The amino acid sequences of all these cyanogen bromide peptides were determined by a combination of tryptic digestion, carboxypeptidase digestion and manual Edman degradation. The large peptides were hydrolyzed with trypsin after maleylation or treatment with 1,2-cyclohexanedione. These cyanogen bromide peptides were aligned by homology with the corresponding cyanogen bromide peptides from pig heart isozyme. The polypeptide chain has 401 amino acid residues and a calculated molecular weight of 44,358. Comparison showed that 24 of the 401 residues of mitochondrial glutamic oxaloacetic transaminase from rat liver are different from those of pig heart isozyme and that homology between the two isozymes is 94%.
 ...
PMID:The complete amino acid sequence of mitochondrial glutamic oxaloacetic transaminase from rat liver. 730 4

1. Above pH 4.3 the outer surface of thylakoid membranes isolated from pea chloroplasts is negatively charged but below this value it carries an excess of positive charge. 2. Previously the excess negative charge has been attributed to the carboxyl groups of glutamic and aspartic acid residues (Nakatani, H.Y., Barber, J. and Forrester, J.A. (1978), Biochim. Biophys. Acta 504, 215-225) and in this paper it is argued from experiments involving treatments with 1,2-cyclohexanedione that the positive charges are partly due to the guanidino group of arginine. 3. The electrophoretic mobility of granal (enriched in chlorophyll b and PS II activity) and stromal (enriched in PS I activity) lamellae isolated by the French Press technique were found to be the same. 4. Treatment of the pea thylakoids with trypsin or pronase, sufficient to inhibit the salt induced chlorophyll fluorescence changes, increased their electrophoretic mobility indicating that additional negative charges had been exposed at the surface. 5. Polylysine treatment also inhibited the salt induced chlorophyll fluorescence changes but unlike trypsin and pronase, decreased the net negative charge on the surface. 6. The isoelectric point defined as the pH which gave zero electrophoretic mobility (about 4.3) was independent of the nature of the cations in the suspending medium (monovalent vs. divalent).
 ...
PMID:Further studies of the thylakoid membrane surface charges by particle electrophoresis. 738 17

The complete primary structure of the major component myoglobin from the goose-beaked whale, Ziphius cavirostris, was determined by specific cleavage of the protein to obtain large peptides which are readily degraded by the automatic sequencer. Over 80% of the amino acid sequence was established from the three peptides resulting from the cleavage of the apomyoglobin at its two methionine residues with cyanogen bromide along with the four peptides resulting from the cleavage with trypsin of the citraconylated apomyoglobin at its three arginine residues. Further digestion of the central cyanogen bromide peptide with S. aureus strain V8 protease and the 1,2-cyclohexanedione-treated central cyanogen bromide peptide with trypsin enabled the determination of the remainder of the covalent structure. This myoglobin differs from the cetacean myoglobins determined to date at 12 to 17 positions. These large sequence differences reflect the distant taxonomic relationships between the goose-beaked whale and the other species of Cetacea the myoglobin sequences of which have previously been determined.
 ...
PMID:Complete amino acid sequence of the major component myoglobin from the goose-beaked whale, Ziphius cavirostris. 743 58

Chemical modification studies have been carried out on the sea anemone polypeptide anthopleurin-A in order to clarify the role of Arg-14 in its cardiac stimulatory activity. Reaction with 1,2-cyclohexanedione at 37 degrees C produced a range of protein products, including some with amino group modifications. These side-reactions were eliminated by prior citraconylation of the amino groups, which, following reaction with cyclohexanedione, could be reversed under conditions which preserved the cyclohexanedione adduct. Citraconylation of the three amino groups, one from the N-terminus and two from Lys-37 and Lys-48, destroyed the cardiac stimulatory activity of the molecule, but this was fully recoverable upon reversal of this reaction. It appears that one or more of the amino groups is essential for activity. Anthopleurin-A contains only one arginine residue, and this was confirmed as the site of modification by cyclohexanedione by showing that the product was refractory to proteolysis by trypsin, which normally cleaves the molecule at this residue. The positive inotropic activity of the cyclohexanedione adduct on isolated guinea-pig atria was identical to that of unmodified anthopleurin-A, indicating that the side-chain of Arg-14 is not required for cardiotonic activity.
 ...
PMID:Chemical modification of cationic groups in the polypeptide cardiac stimulant anthopleurin-A. 759 22

Sulfate derived from the degradation of macromolecules is released from lysosomes via a carrier mediated process. In order to further characterize this process, recognized inhibitors of the erythrocyte band 3 anion transporter were examined for their effects on the lysosomal system. Studies with band 3 transport site inhibitors such as DIDS, SITS and phenylglyoxal indicated that, similar to the case for the band 3 protein, the lysosomal transporter has critical lysine and arginine residues. Band 3 translocation pathway or channel blocking inhibitors had mixed effects on the lysosomal system. 1,2-Cyclohexanedione, which covalently modifies a band 3 arginine residue distinct from that modified by phenylglyoxal, inhibited lysosomal sulfate transport. In contrast, the potent band 3 inhibitor dipyridamole had no effect on lysosomal sulfate transport indicating that there are some structural differences between the erythrocyte and lysosomal anion transporters. The band 3 translocation inhibitors niflumic acid and dinitrofluorobenzene were both effective inhibitors of the lysosomal system. Cupric ion inhibited sulfate transport while Ca2+, Co2+, Mg2+, Mn2+, and Zn2+ had no inhibitory effects. Exposure of intact lysosomes to trypsin largely ablated transport of sulfate. This information should be useful in efforts to further elucidate the structure and function of the lysosomal sulfate transporter.
 ...
PMID:Lysosomal sulfate transport: inhibitor studies. 771 10

A protein modification method has been developed for the production of human big endothelin (ET)-1. Production of a large quantity of big ET-1 by the method described here is expected to facilitate future experiments such as X-ray crystallography and nuclear magnetic resonance studies, aimed at understanding the tertiary structure of big ET-1 and its dynamics. The plasmid pETB-50 used for the synthesis carries the gene for a fusion protein consisting of 34-amino acid (aa) residues of an N-terminal portion of beta-galactosidase and the 38-aa residues of big ET-1. The fusion protein ETB-50P contains an arginine residue in the big ET-1 portion at its second C-terminal site and three lysine residues including the C-terminal site in the beta-galactosidase portion, all of which are susceptible to trypsin. Tryptic digestion of the fusion protein quantitatively produced big ET-1 (1-37), which is depleted in the C-terminal serine. However, a treatment of the fusion protein with 1,2-cyclohexanedione prior to tryptic digestion gave full-length big ET-1 with N7, -N8-(1,2-dihydroxycyclohex-1,2- ylene)-arginine. This modification was reversed to the intact arginine residue when the modified big ET-1 was incubated in 0.5 M TRIS-HCl buffer, pH 8.0. Consequently, a combination of such a reversible protein modification and tryptic digestion gave 1.74 mg of recombinant big ET-1 from 2.01 of culture broth. The procedure described here may be applied to produce other arginine-containing peptides from fusion proteins.
 ...
PMID:High-yield production of human big endothelin-1 by a combination of chemical modification and proteolysis of a fusion protein in Escherichia coli. 776 64


<< Previous 1 2 3