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G Salvesen

Publications and source records attributed to G Salvesen.

46 records · Page 3Linked to original sources
PubMed1988

Human kininogens.

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Chromatography, Affinity↗
PubMed1988

Molecular cloning of human neutrophil elastase.

The human U937 cell line, a promonocyte-like leukemic line, has been shown to synthesize neutrophil elastase (Senior, R. M. et al. (1982) J. Clin. Invest. 69, 384-393). We have constructed a lambda gt11 cDNA library using RNA from this cell line and isolated a clone encoding part of the protein sequence of neutrophil elastase. Nucleotide sequencing indicates the clone encodes residues 108-243 (standard chymotrypsinogen numbering) of the enzyme, plus a hitherto unsuspected 20 residue-C-terminal extension of unusual structure. Comparison of the nucleotide and amino-acid sequence of elastase with the recently reported sequence of medullasin, a proteinase isolated from bone marrow, suggests they are identical.

Amino Acid Sequence↗
PubMed1987

Rapid isolation of human kininogens.

A rapid, two-step procedure is described for the isolation of both "high molecular weight" (H-) and "low molecular weight" (L-) plasma kininogens from a single sample of plasma. Affinity chromatography on carboxymethyl-papain-Sepharose is used, together with high-resolution anion exchange chromatography.

Blood Coagulation↗
PubMed1987

Molecular cloning of human cathepsin G: structural similarity to mast cell and cytotoxic T lymphocyte proteinases.

Human cathepsin G is a serine proteinase with chymotrypsin-like specificity found in both polymorphonuclear leukocytes (neutrophils) and the U937 leukemic cell line. Utilizing RNA from the latter, we have constructed a cDNA library in lambda gt11 and isolated a clone which apparently codes for the complete amino acid sequence of this enzyme. Analysis of the sequence reveals homology with rat mast cell proteinase II (47%) but a greater degree of identity (56%) with a product of activated mouse cytotoxic T lymphocytes. The close relationship between the three proteins indicates similarities in substrate specificity and in biosynthesis which we predict involves removal of a two amino acid activation peptide during or just before packaging into their respective storage granules.

Amino Acid Sequence↗
PubMed1986

Isolation of six cysteine proteinase inhibitors from human urine. Their physicochemical and enzyme kinetic properties and concentrations in biological fluids.

Six cysteine proteinase inhibitors were isolated from human urine by affinity chromatography on insolubilized carboxymethylpapain followed by ion-exchange chromatography and immunosorption. Physicochemical and immunochemical measurements identified one as cystatin A, one as cystatin B, one as cystatin C, one as cystatin S, and one as low molecular weight kininogen. The sixth inhibitor displayed immunochemical cross-reactivity with salivary cystatin S but had a different pI (6.85 versus 4.68) and a different (blocked) N-terminal amino acid. This inhibitor was tentatively designated cystatin SU. The isolated inhibitors accounted for nearly all of the cysteine proteinase inhibitory activity of the urinary pool used as starting material. The enzyme inhibitory properties of the inhibitors were investigated by measuring inhibition and rate constants for their interactions with papain and human cathepsin B. Antisera raised against the inhibitors were used in immunochemical determinations of their concentrations in several biological fluids. The combined enzyme kinetic and concentration data showed that several of the inhibitors have the capacity to play physiologically important roles as cysteine proteinase inhibitors in many biological fluids. Cystatin C had the highest molar concentration of the inhibitors in seminal plasma, cerebrospinal fluid, and milk; cystatin S in saliva and tears; and kininogen in blood plasma, synovial fluid, and amniotic fluid.

Body Fluids↗
PubMed1986

Human low-Mr kininogen contains three copies of a cystatin sequence that are divergent in structure and in inhibitory activity for cysteine proteinases.

We point out that human low-Mr kininogen contains three cystatin-like sequences, rather than two, as had previously been thought. The protein was purified by affinity chromatography on carboxymethyl-papain-Sepharose, and subjected to limited proteolysis by trypsin and chymotrypsin. Fragments were isolated, and three corresponding to the individual cystatin-like domains were identified. By comparison with the known amino acid sequence of the protein they were numbered 1 to 3 from the N-terminus. Domain 1 was not found to have any inhibitory activity for cysteine proteinases, which is consistent with the absence of residues that are highly conserved in inhibitors of the cystatin superfamily, and have previously been suggested to be essential for activity. Domain 2 was a good inhibitor of chicken calpain, and also papain and cathepsin L. Domain 3 showed negligible inhibition of calpain, but inhibited papain and cathepsin L strongly. The probable arrangement of disulphide bonds in the heavy chain of low-Mr kininogen is deduced from the homology with the cystatins and other evidence contained in the present paper.

Amino Acid Sequence↗
PubMed1983

Control of coagulation and fibrinolysis by plasma proteinase inhibitors.

The control of coagulation and fibrinolytic events appears to be primarily due to four plasma proteinase inhibitors, antithrombin III, C-1-esterase inhibitor, alpha-2-antiplasmin, and alpha-2-macroglobulin. Results to date indicate that antithrombin III controls the activity of both thrombin and Factor Xa, C-1-esterase inhibitor controls kallikrein and probably activated Hageman Factor (Factor XIIa), and alpha-2-antiplasmin controls plasmin activity. The role of alpha-2-macroglobulin is not clear since it does not appear to be a primary inhibitor of any of the above enzymes. However, it is probable that it serves two functions, first as a "transfer" agent for the rapid removal of proteinases from the circulation which have been first bound by antithrombin III, C-1-esterase inhibitor, or alpha-2-antiplasmin. The second function is probably that of a back-up inhibitor when the levels of the three important controlling plasma proteins become low. The role of other plasma inhibitors such as alpha-1-proteinase inhibitor, alpha-1-antichymotrypsin, and the inter-alpha-trypsin inhibitor in coagulation and fibrinolysis would appear to be minor since these proteins either do not inactivate enzymes involved in these systems or do so at a rate too slow to be of biological significance.

Antifibrinolytic Agents↗