Biomaterial Database

Purification to homogeneity of the human skin chymotryptic proteinase "chymase". 1986

L A Johnson, and K E Moon, and M Eisenberg

The chymotrypic proteinase "chymase" has been purified to apparent homogeneity from human skin. Our procedure differs from previously published partial purifications in that it does not involve affinity chromatography, most of the steps are carried out in 2 M KCl which stabilizes the enzyme, detergent is used to protect the enzyme in low-ionic-strength media, and troublesome concentration steps are avoided by using very small columns of high-capacity exchangers. The high-salt skin extract is applied successively to columns of hydroxyapatite, copper chelate Sepharose, and Sephadex G-100 in 2 M KCl. After dialysis against a zwitterionic detergent, the enzyme is adsorbed onto a 0.4-ml column of CM-Sepharose. An alkaline wash removes the remaining contaminants from the highly cationic enzyme, which is then eluted with 1 M KCl in a final volume of 2 ml. Sodium dodecyl sulfate electrophoresis reveals a single diffuse band of Mr 30,000. Recoveries range from 20 to 40% with yields of 0.2 to 0.4 mg of enzyme from 200 g of skin. Specific activities vary from 600 to 1400 units/mg for the hydrolysis of acetyltyrosine ethyl ester.

UI	MeSH Term	Description	Entries
D010450	Endopeptidases	A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.	Endopeptidase,Peptide Peptidohydrolases
D002845	Chromatography	Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.	Chromatographies
D003300	Copper	A heavy metal trace element with the atomic symbol Cu, atomic number 29, and atomic weight 63.55.	Copper-63,Copper 63
D003911	Dextrans	A group of glucose polymers made by certain bacteria. Dextrans are used therapeutically as plasma volume expanders and anticoagulants. They are also commonly used in biological experimentation and in industry for a wide variety of purposes.	Dextran,Dextran 40,Dextran 40000,Dextran 70,Dextran 75,Dextran 80,Dextran B-1355,Dextran B-1355-S,Dextran B1355,Dextran B512,Dextran Derivatives,Dextran M 70,Dextran T 70,Dextran T-40,Dextran T-500,Hemodex,Hyskon,Infukoll,Macrodex,Polyglucin,Promit,Rheodextran,Rheoisodex,Rheomacrodex,Rheopolyglucin,Rondex,Saviosol,Dextran B 1355,Dextran B 1355 S,Dextran T 40,Dextran T 500
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012685	Sepharose		Agarose,Sepharose 4B,Sepharose C1 4B,4B, Sepharose C1,C1 4B, Sepharose
D012697	Serine Endopeptidases	Any member of the group of ENDOPEPTIDASES containing at the active site a serine residue involved in catalysis.	Serine Endopeptidase,Endopeptidase, Serine,Endopeptidases, Serine
D012867	Skin	The outer covering of the body that protects it from the environment. It is composed of the DERMIS and the EPIDERMIS.
D053818	Chymases	A family of neutral serine proteases with CHYMOTRYPSIN-like activity. Chymases are primarily found in the SECRETORY GRANULES of MAST CELLS and are released during mast cell degranulation.	Chymase,Chymase-1,Chymotrypsin-Like Protease,Chymotrypsin-Like Proteases,Chymase 1,Chymotrypsin Like Protease,Chymotrypsin Like Proteases