Hydrolysis of alanine oligomers and of elastin by P. aeruginosa proteinases and thermolysin.
The hydrolysis of alanine oligomers by P. aeruginosa proteinases, thermolysin and porcine pancreatic elastase was studied. The concentrations of substrates and cleavage products were determined using reverse phase high pressure liquid chromatography. Tetraalanine was the shortest oligomer for which we could demonstrate hydrolysis by all the proteinases, except for porcine pancreatic elastase which only significantly hydrolyzed peptides longer than hexaalanine. Porcine pancreatic elastase hydrolyzes hexaalanine at a single site, whereas the other enzymes may split it either into two trialanine molecules, or into di- and tetraalanine, the latter being further cleavable to dialanine. A kinetic model based on first-order kinetic rate constants is proposed and the individual constants determined. Although P. aeruginosa elastase and thermolysin are closely similar in structure, they have shown a marked difference in their hydrolysis of either elastin or tetraalanine. Elastolytic activity of thermolysin was higher than that of elastase but tetraalanine was hydrolyzed more slowly by thermolysin.
Saulnier
J
J
Biochimie Analytique, I.C.B.M.C., Université Lyon I, Villeurbanne, France.
Thevenon
F
F
Besson
C
C
Duclos
M C
MC
Wallach
J M
JM
D000409:Alanine / Q000378:metabolism*; D001426:Bacterial Proteins; D002851:Chromatography, High Pressure Liquid; D004549:Elastin / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013820:Thermolysin / Q000378:metabolism*
D001426:Bacterial Proteins; D009842:Oligopeptides; D004549:Elastin; D010450:Endopeptidases; D008666:Metalloendopeptidases; C043859:pseudolysin, Pseudomonas aeruginosa; D013820:Thermolysin; D000409:Alanine