Apparent suppression of MMP-9 activity by GD1a as determined by gelatin zymography

Dan Hu, Xuan Tan, Toshinori Sato, Sadako Yamagata, Tatsuya Yamagata

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Gelatin zymography is widely used to detect and evaluate matrix metalloproteinase-9 (MMP-9) activity. MMP-9 transcription was previously shown to be negatively regulated by ganglioside GD1a [D. Hu, Z. Man, T. Xuan, P. Wang, T. Takaku, S. Hyuga, X.S. Yao, T. Sato, S. Yamagata, T. Yamagata, Ganglioside GD1a regulation of matrix metalloproteinase-9 (MMP-9) expression in mouse FBJ cell Lines: GD1a suppression of MMP-9 expression stimulated by PI3K-Akt and p38 though not by the Erk signaling pathway, 2006, submitted for publication.]. Zymography of MMP-9 of FBJ-M5 cells preincubated with GD1a indicated a greater decrease in activity than expected from mRNA suppression. Incubation of conditioned medium containing MMP-9 with GD1a caused MMP-9 activity to decrease. Examination was thus made to confirm that MMP-9 activity is actually suppressed and/or MMP-9 protein undergoes degradation by GD1a. GD1a was found to have no effect on MMP-9 activity and Western blots indicated GD1a not to diminish MMP-9 during electrophoresis under reducing conditions. GD1a appeared to mediate the binding of a portion of MMP-9 with certain molecules, with consequently greater molecular mass on the gel, to cause decrease in the activity of MMP-9 at the site where it would normally appear. Caution should be used in doing gelatin zymography since molecules other than GD1a may similarly work, causing decrease in MMP-9 activity in zymography.

Original languageEnglish
Pages (from-to)426-431
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume349
Issue number1
DOIs
Publication statusPublished - 2006 Oct 13

Fingerprint

Matrix Metalloproteinase 9
Gelatin
Molecules
Molecular mass
Transcription
Conditioned Culture Medium
Electrophoresis
Phosphatidylinositol 3-Kinases
Proteolysis
Publications

Keywords

  • Ganglioside
  • GD1a
  • Metastasis
  • MMP-2
  • MMP-9
  • Zymography

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Apparent suppression of MMP-9 activity by GD1a as determined by gelatin zymography. / Hu, Dan; Tan, Xuan; Sato, Toshinori; Yamagata, Sadako; Yamagata, Tatsuya.

In: Biochemical and Biophysical Research Communications, Vol. 349, No. 1, 13.10.2006, p. 426-431.

Research output: Contribution to journalArticle

Hu, Dan ; Tan, Xuan ; Sato, Toshinori ; Yamagata, Sadako ; Yamagata, Tatsuya. / Apparent suppression of MMP-9 activity by GD1a as determined by gelatin zymography. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 349, No. 1. pp. 426-431.
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abstract = "Gelatin zymography is widely used to detect and evaluate matrix metalloproteinase-9 (MMP-9) activity. MMP-9 transcription was previously shown to be negatively regulated by ganglioside GD1a [D. Hu, Z. Man, T. Xuan, P. Wang, T. Takaku, S. Hyuga, X.S. Yao, T. Sato, S. Yamagata, T. Yamagata, Ganglioside GD1a regulation of matrix metalloproteinase-9 (MMP-9) expression in mouse FBJ cell Lines: GD1a suppression of MMP-9 expression stimulated by PI3K-Akt and p38 though not by the Erk signaling pathway, 2006, submitted for publication.]. Zymography of MMP-9 of FBJ-M5 cells preincubated with GD1a indicated a greater decrease in activity than expected from mRNA suppression. Incubation of conditioned medium containing MMP-9 with GD1a caused MMP-9 activity to decrease. Examination was thus made to confirm that MMP-9 activity is actually suppressed and/or MMP-9 protein undergoes degradation by GD1a. GD1a was found to have no effect on MMP-9 activity and Western blots indicated GD1a not to diminish MMP-9 during electrophoresis under reducing conditions. GD1a appeared to mediate the binding of a portion of MMP-9 with certain molecules, with consequently greater molecular mass on the gel, to cause decrease in the activity of MMP-9 at the site where it would normally appear. Caution should be used in doing gelatin zymography since molecules other than GD1a may similarly work, causing decrease in MMP-9 activity in zymography.",
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