Reactivity of antibodies against 10-amino acid residue and pro-domain of stromelysin-3.
Stromelysin-3 (ST3) has two highly conserved domains in the pro-domain. In particular, an unusual 10-amino acid residue sandwiched between the pro-domain and the catalytic domain of ST3 exists in ST3 but not in other matrix metalloproteinases (MMPs). To specifically detect ST3 expression in human tumors, we have made two kinds of ST3-specific polyclonal antibodies. One was raised against the synthetic 10-amino acid residue (88GLSARNRQKR97) specific to ST3, and the other against recombinant ST3 pro-domain (62APATQEAPRPASSLRPPRCGVPDPSDGLSARNRQKR97) containing the decapeptide and PRCGVPD sequence obtained by expression in Escherichia coli. Two protein species, 59 kDa and 45 kDa which were consistent with those expected for pro-ST3 and the mature form of ST3, were specifically detected in 100-fold concentrated conditioned media of fetal lung fibroblast by Western blot analysis. Immunohistochemical staining indicated that in infiltrating ductal breast carcinoma and squamous cell carcinoma of the uterine cervix, reactivity of those antibodies was found not only in fibroblastic cells surrounding cancer cells but also in neoplastic cells. However, reactivity of two ST3 antibodies was inhibited by excess of the synthetic peptide (10-amino acid residue) not only in fibroblastic cells but also in neoplastic cells. These findings suggest that antibodies against the ST3 specific region may cross react with the recently known membrane type-metalloproteinase (MT-MMP), which have RXKR sequences between the pro- and catalytic domain.
Lyu
M A
MA
Department of Microbiology and Cancer Research Center, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea.
Park
I A
IA
Hong
S I
SI
Kim
B J
BJ
Park
I K
IK
Goh
Y J
YJ
Kim
S J
SJ
Hwang
E S
ES
Kook
Y H
YH
D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D018270:Carcinoma, Ductal, Breast / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism; D003429:Cross Reactions; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D053507:Matrix Metalloproteinase 11 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D009842:Oligopeptides / Q000276:immunology / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014594:Uterine Neoplasms / Q000378:metabolism
D000906:Antibodies; D009842:Oligopeptides; C511771:MMP11 protein, human; D053507:Matrix Metalloproteinase 11