Characterization of 58-kilodalton human neutrophil collagenase: Comparison with human fibroblast collagenase

Journal Article · Thu Nov 01 00:00:00 EST 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00499a008· OSTI ID:5614822

Mallya, S K; Mookhtiar, K A; Gao, Y; Dioszegi, M; Wart, H E.V. ^[1]; Brew, K ^[2]; Birkedal-Hansen, H ^[3]

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1. Florida State Univ., Tallahassee, FL (USA)
2. Univ. of Miami Medical School, FL (USA)
3. Univ. of Alabama School of Dentistry, Birmingham (USA)

A series of experiments has been carried out to characterize 58-kDa human neutrophil collagenase (HNC) and compare it with human fibroblast collagenase (HFC). N-Terminal sequencing of latent and spontaneously activated HNC shows that it is a distinct collagenase that is homologous to HFC and other members of the matrix metalloproteinase gene family. Activation occurs autolytically by hydrolysis of an M-L bond at a locus homologous to the Q{sub 80}-F{sub 81}-V{sub 82}-L{sub 83} autolytic activation site of HFC. This releases a 16-residue propeptide believed to contain the cysteine switch residue required for latency. Polyclonal antibody raised against HNC cross-reacts with HFC but with none of the other major human matrix metalloproteinases examined. Treatment of HNC with endoglycosidase F or N-glycosidase F indicates that it is glycosylated at multiple sites. The deglycosylated latent and spontaneously activated enzymes have molecular weights of approximately 44K and 42K, respectively. Differences in the carbohydrate processing of HFC and HNC may determine why HFC is a secreted protein while HNC is stored in intracellular granules. The kinetic parameters K{sub cat} and K{sub M} for the hydrolysis of the interstitial collagen types I, II, and III in solution by both collagenases have been determined. The rates of hydrolysis of these peptides vary very little, indicating that it is the collagen conformation at the cleavage site and not the sequence specificity of the collagenases that determines their collagen specificities.

• OSTI ID: 5614822
• Journal Information: Biochemistry; (USA), Vol. 29:47; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
COLLAGEN
AMINO ACID SEQUENCE
BIOCHEMISTRY
ELECTROPHORESIS
ENZYMATIC HYDROLYSIS
FIBROBLASTS
NEUTROPHILS
PEPTIDE HYDROLASES
ANIMAL CELLS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CHEMICAL REACTIONS
CHEMISTRY
CONNECTIVE TISSUE CELLS
DECOMPOSITION
ENZYMES
HYDROLASES
HYDROLYSIS
LEUKOCYTES
LYSIS
MATERIALS
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
PROTEINS
SCLEROPROTEINS
SOLVOLYSIS
SOMATIC CELLS
550201* - Biochemistry- Tracer Techniques