Enzymatic and molecular biochemical characterizations of human neutrophil elastases and a cathepsin G-like enzyme.
Human neutrophil elastase (HNE, IEC 3. 4. 21. 37) is a causative factor of inflammatory diseases, including emphysema and rheumatoid arthritis. Enzymatic characterization is important for the development of new drugs involved in the regulation of this enzyme. In this study, we investigated the enzymatic and biochemical properties of five different elastolytic enzymes, with a molecular mass between 24 kDa and 72 kDa. Three elastases, molecular masses of 27, 29, 31 kDa, might be elastase isozymes that have the same NH2-terminal amino acid sequences of Ile-Val-Gly-Gly-Arg-Arg-Ala. The 24-kDa enzyme, which showed the identical NH2-terminal amino acid sequences to elastase, was a degraded fragment of native elastase. The elastolytic activity was conserved at the 6/7 domain of the NH2-terminal region. The inhibitory characteristics of PMSF, DipF were the same as those of native elastases. The 72-kDa molecule, which showed elastolytic activity, might be a trimer formed between native elastases (31 kDa and 29 kDa) and a cathepsin G-like enzyme, which did not show elastolytic activity but enhanced the elastolytic activity of neutrophil elastase. Although this cathepsin G-like enzyme showed weak cathepsin G activity, it has distinguishable NH2-terminal sequences of Ile-Val-Gly-Gly-Ser-Arg-Ala- from those of elastase or cathepsin G. The potentiation of elastolytic activity could be a result of the trimerization of native elastase with a cathepsin G-like enzyme, and was then weakly inhibited by serine protease inhibitors, such as PMSF, DipF. Therefore, we suggest the cathepsin G-like enzyme to be a novel enzyme, which has an important role in the development of inflammation.
Kim
W M
WM
Department of Pharmacology, Kosin University Medical College, Pusan, Korea. kwm@ns.kosinmed.or.kr
Kang
K
K
D000595:Amino Acid Sequence; D056649:Cathepsin G; D002403:Cathepsins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D018360:Crystallography, X-Ray; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D007700:Kinetics; D019272:Leukocyte Elastase / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D008958:Models, Molecular; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology*; D010446:Peptide Fragments / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012697:Serine Endopeptidases
D007527:Isoenzymes; D010446:Peptide Fragments; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D019272:Leukocyte Elastase