The cleavage of bovine collagen I by neutrophil collagenase MMP-8 has been followed at pH 7.4, 37 degreesC. The behavior of the whole enzyme molecule (whMMP-8), displaying both the catalytic domain and the hemopexin-like domain, has been compared under the same experimental conditions with that of the catalytic domain only. The main observation is that whMMP-8 cleaves bovine collagen I only at a single specific site, as already reported by many others (Mallya, S. K., Mookhtiar, K. A., Gao, Y., Brew, K., Dioszegi, M., Birkedal-Hansen, H., and van Wart, H. E. (1990) Biochemistry 29, 10628-10634; Knauper, V., Osthues, A., DeClerk, Y. A., Langley, K. A, Blaser, J., and Tschesche, H. (1993) Biochem. J. 291, 847854; Marini, S., Fasciglione, G. F., De Sanctis, G., D'Alessio, S., Politi, V., and Coletta, M. (2000) J. Biol. Chem. 275, 18657-18663), whereas the catalytic domain lacks this specificity and cleaves the collagen molecule at multiple sites. Furthermore, a meaningful difference is observed for the cleavage features displayed by two forms of the catalytic domain, which differ for the N terminus resulting from the activation process (i.e. the former Met(80)of the proenzyme (MetMMP-8) and the former Phe(79) of the proenzyme (PheMMP-8)). Thus, the PheMMP-8 species is characterized by a much faster k(cat)/K-m, fully attributable to a lower K-m, suggesting that the conformation of the catalytic domain, induced by the insertion of this N-terminal residue in a specific pocket (Reinemer, P., Grams, F., Huber, R., Kleine, T., Schnierer, S., Piper, M., Tschesche, H., and Bode, W. (1994) FEBS Lett. 338, 227-233), brings about a better, although less discriminatory, recognition process of cleavage site(s) on bovine collagen I.

Gioia, M., Fasciglione, G., Marini, S., D'Alessio, S., De Sanctis, G., Diekmann, O., et al. (2002). Modulation of the catalytic activity of neutrophil collagenase MMP-8 on bovine collagen I - Role of the activation cleavage and of the hemopexin-like domain. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 277(26), 23123-23130 [10.1074/jbc.M110873200].

Modulation of the catalytic activity of neutrophil collagenase MMP-8 on bovine collagen I - Role of the activation cleavage and of the hemopexin-like domain

GIOIA, MAGDA;FASCIGLIONE, GIOVANNI;MARINI, STEFANO;COLETTA, MASSIMILIANO
2002-06

Abstract

The cleavage of bovine collagen I by neutrophil collagenase MMP-8 has been followed at pH 7.4, 37 degreesC. The behavior of the whole enzyme molecule (whMMP-8), displaying both the catalytic domain and the hemopexin-like domain, has been compared under the same experimental conditions with that of the catalytic domain only. The main observation is that whMMP-8 cleaves bovine collagen I only at a single specific site, as already reported by many others (Mallya, S. K., Mookhtiar, K. A., Gao, Y., Brew, K., Dioszegi, M., Birkedal-Hansen, H., and van Wart, H. E. (1990) Biochemistry 29, 10628-10634; Knauper, V., Osthues, A., DeClerk, Y. A., Langley, K. A, Blaser, J., and Tschesche, H. (1993) Biochem. J. 291, 847854; Marini, S., Fasciglione, G. F., De Sanctis, G., D'Alessio, S., Politi, V., and Coletta, M. (2000) J. Biol. Chem. 275, 18657-18663), whereas the catalytic domain lacks this specificity and cleaves the collagen molecule at multiple sites. Furthermore, a meaningful difference is observed for the cleavage features displayed by two forms of the catalytic domain, which differ for the N terminus resulting from the activation process (i.e. the former Met(80)of the proenzyme (MetMMP-8) and the former Phe(79) of the proenzyme (PheMMP-8)). Thus, the PheMMP-8 species is characterized by a much faster k(cat)/K-m, fully attributable to a lower K-m, suggesting that the conformation of the catalytic domain, induced by the insertion of this N-terminal residue in a specific pocket (Reinemer, P., Grams, F., Huber, R., Kleine, T., Schnierer, S., Piper, M., Tschesche, H., and Bode, W. (1994) FEBS Lett. 338, 227-233), brings about a better, although less discriminatory, recognition process of cleavage site(s) on bovine collagen I.
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/10
English
Con Impact Factor ISI
Activation analysis; Catalysis; Collagen; Conformations; Enzymes; Cleavage sites; Biochemistry; collagen type 1; hemopexin; neutrophil collagenase; amino terminal sequence; article; catalysis; cattle; enzyme activation; enzyme activity; enzyme analysis; enzyme specificity; nonhuman; priority journal; protein domain; Animals; Catalytic Domain; Cattle; Collagen Type I; Enzyme Activation; Matrix Metalloproteinase 8; Neutrophils; Bos taurus; Bovinae; insertion sequences
Gioia, M., Fasciglione, G., Marini, S., D'Alessio, S., De Sanctis, G., Diekmann, O., et al. (2002). Modulation of the catalytic activity of neutrophil collagenase MMP-8 on bovine collagen I - Role of the activation cleavage and of the hemopexin-like domain. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 277(26), 23123-23130 [10.1074/jbc.M110873200].
Gioia, M; Fasciglione, G; Marini, S; D'Alessio, S; De Sanctis, G; Diekmann, O; Pieper, M; Politi, V; Tschesche, H; Coletta, M
Articolo su rivista
File in questo prodotto:
File Dimensione Formato  
JBC2002MMP8.pdf

accesso aperto

Dimensione 269.02 kB
Formato Adobe PDF
269.02 kB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2108/53099
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact