Matrix metalloproteinase activities of turkey (Meleagris gallopavo) bile.
The bile from turkey (Meleagris gallopavo) gall bladders was found to contain substantial matrix metalloproteinase (MMP) activities using gelatin, collagen, and casein substrate zymography, [3H]labeled collagen degradation assays, and gelatin-agarose affinity purification. Five major bands corresponding to approximate M(w) of 64, 60, 46, 40 and 36 kDa showed gelatinolytic activities. On incubation with p-aminophenylmercuric acetate or thimerosal, the densities of both the 64- and 46-kDa bands decreased with increasing intensities of the 60- and 40-kDa bands. Both the 64- and 60-kDa bands showed collagenolytic activities whereas the caseinolytic activities appeared as diffuse bands corresponding to M(w) of approximately 60, 40 and 36 kDa. Using [3H]collagen as substrate, the bile enzymes showed both a time and concentration-dependent degradation, which could be inhibited by the MMP inhibitors such as EDTA, phenanthroline, and N-[(2R)-2-(hydroxyamido carbonylmethyl)-4-methylpentanonyl]-L-tryptophan methylamide, but not by serine and cysteine protease inhibitors like trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, phenylmethylsulfonyl fluoride or leupeptin. Both 60- and the 40-kDa gelatinolytic bands showed affinity adsorption to a gelatin-agarose matrix. The physiological roles of bile MMPs are not clear, but their involvement in the digestive functions of birds are likely.
Rath
N C
NC
USDA, ARS, PPPSRU, Poultry Science Center, University of Arkansas, Fayetteville, AR 72701, USA. nrath@uark.edu
Huff
W E
WE
Huff
G R
GR
Balog
J M
JM
Xie
H
H
D000818:Animals; D001646:Bile / Q000201:enzymology*; D015153:Blotting, Western; D002645:Chickens; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases / Q000032:analysis / Q000276:immunology; D005780:Gelatin / Q000737:chemistry; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D012685:Sepharose / Q000737:chemistry; D014422:Turkeys
D005780:Gelatin; D003094:Collagen; D012685:Sepharose; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases