Biochemical properties of metalloproteinases from the hemolymph of the mussel Mytilus galloprovincialis Lam.
The expression of matrix metalloproteinases (MMP) with gelatinase activity was found in the whole hemolymph of the marine mussel Mytilus galloprovincialis Lam. Cleavage activity was specific for gelatin; very little activity towards human type-IV collagen, and no activity for cold fish gelatin, casein or bovine serum albumin were detected. EDTA and 1,10-phenanthroline were inhibitory, suggesting that mussel MMPs require divalent cations for their proteolytic activity; in fact, the presence of exogenously added divalent ions significantly protected the MMPs from inhibition. No inhibition was detected with serine or cysteine proteinase inhibitors. The specific vertebrate inhibitors as well as the classical vertebrate activator of MMPs were without effect, whereas sulphydryl reducing agents had a strong inhibitory effect. Mussel MMPs showed an exponential curve of thermal-dependent decay that was not protected by the presence of metal ions. Overall the results indicate both similarities and differences between invertebrate and vertebrate gelatinases, providing information for understanding the biological role of these ancient proteinases.
Mannello
F
F
Istituto di Istologia ed Analisi di Laboratorio, Facoltà di Scienze Matematiche, Fisiche e Naturali, Libera Università degli Studi, Via E. Zeppi, 61029 Urbino (PU), Italy. mannello@uniurb.it
Canesi
L
L
Gazzanelli
G
G
Gallo
G
G
D000818:Animals; D049872:Bivalvia / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002850:Chromatography, Gel; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005780:Gelatin / Q000378:metabolism; D006458:Hemolymph / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D010618:Phenanthrolines / Q000494:pharmacology; D013379:Substrate Specificity; D015032:Zinc / Q000494:pharmacology
D004791:Enzyme Inhibitors; D010618:Phenanthrolines; D005780:Gelatin; D004492:Edetic Acid; D008666:Metalloendopeptidases; D008274:Magnesium; D015032:Zinc; D002118:Calcium