Complexes of gelatinases and tissue inhibitor of metalloproteinases in human seminal plasma.
Previously reported data have indicated the existence of two kinds of matrix metalloproteinases (MMP-2 and MMP-9) in human seminal plasma (Shimokawa et al, 2002). Here we report the existence of complexes of gelatinases and tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 in human seminal plasma. After the seminal plasma supernatant was separated on a gel-filtration column chromatography of GCL-2000-sf-cellulofine. Western blot analysis showed these proteins were recognized by two antibodies to TIMP-1 and TIMP-2, but not to TIMP-3 or TIMP-4. These bands were consistent with standard recombinant full-length TIMP-1 and TIMP-2 proteins. These bands had molecular weights of approximately 29 and 21 kd for TIMP-1 and TIMP-2, respectively. These proteins existed as complexes of proMMP-9/TIMP-1, proMMP-2/TIMP-2, MMP-2/TIMP-2, free TIMP-1, and TIMP-2 in human seminal plasma. The partially free TIMPs were degradated by some proteinases in human seminal plasma. These results indicate two kinds of TIMPs (TIMP-1 and TIMP-2) and their complexes with progelatinases in human seminal plasma.
Shimokawa
Ken-Ichi
K
Departments of Functional Bioanalysis and Chemistry of Hygiene, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
Katayama
Masatoki
M
Matsuda
Yoshifumi
Y
Takahashi
Hidenobu
H
Hara
Izumi
I
Sato
Hirohisa
H
D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D012661:Semen / Q000201:enzymology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism*; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism*
D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9