Identification and characterization of the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) in hen egg.
Using zymography and mass spectrometry, we identified for the first time the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) as a complex with TIMP-2 (tissue inhibitor of metalloproteinases) in egg white and yolk. Real-time polymerase chain reaction confirmed that MMP-2 and its inhibitors TIMP-2 and TIMP-3 were expressed all along the oviduct and in the liver of laying hens. We also demonstrated that the processing of pro-MMP-2 into mature MMP-2 by serine proteases does not occur in vivo, although purified pro-MMP-2 undergoes proteolytic maturation by these proteases in vitro. Moreover, the relative pro-MMP-2 activity assessed by gelatin zymography was shown to decrease in egg white during the storage of unfertilized or fertilized eggs. However, the mature form of 62 kDa MMP-2 could not be detected. The fact that MMP-2 is found as a proform in fresh eggs suggests that the activity of this metalloprotease is regulated under specific conditions during embryonic development.
Réhault-Godbert
Sophie
S
INRA, UR83 Recherches Avicoles, Fonction et Regulation des Proteines de l'Oeuf, F-37380 Nouzilly, France. rehault@tours.inra.fr
Gautron
Joël
J
Labas
Valérie
V
Belghazi
Maya
M
Nys
Yves
Y
D000818:Animals; D002642:Chick Embryo / Q000201:enzymology / Q000254:growth & development; D002645:Chickens; D004529:Egg White / Q000032:analysis; D004530:Egg Yolk / Q000201:enzymology; D004531:Eggs / Q000032:analysis*; D004792:Enzyme Precursors / Q000032:analysis*; D005260:Female; D015870:Gene Expression; D020778:Matrix Metalloproteinase 2 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010057:Oviducts / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics
D004792:Enzyme Precursors; D012333:RNA, Messenger; D019717:Tissue Inhibitor of Metalloproteinase-3; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2