Gelatin Zymography Band Interpretation

Youssef Farhat, Written 4/8/13, Last Updated 5/10/13

Here, we summarize possible ways of interpreting individual bands that can be visualized on gelatin zymography based on their molecular weight.  These interpretations have been reported in the papers cited below.  Note that these interpretations need further confirmation with techniques such as Western blots or cutting out the bands and sending them off for sequencing by a proteomics center (let me know if you are aware of other ways to identify them by sending an email to youssef@protocol-place.com).

Molecular Weight (kDa)

Possible Interpretation

240
  • Possible MMP multimer [1]
215
  • MMP-9 dimer associated with 25-kDa microglobulin [1]
~200
  • MMP-9 dimer [2]
180
  • MMP-9 dimer (Chemicon International gelatinase zymography standards, #CC073)
~140-144
  • MMP-2 dimer [3]
125
  • MMP-9 associated with 25-kDa microglobulin [1]
92
  • Pro-MMP-9 [2]
83-85
  • Intracellular precursor, partially glycosylated MMP-9 [2]
  • Intermediate active form of MMP-9 when TIMPs present during APMA activation [2]
82
  • Active MMP-9 [2]
72
  • Pro-MMP-2 [2]
67-68
  • MMP-9 after long incubation with APMA [2]
  • Active MMP-9 [4]
  • Mature MMP-2 formed after AMPA treatment [4]
64
  • Intermediate active form of MMP-2 when TIMPs present during APMA activation [2]
62
  • Active MMP-2 [2]
58
  • Active MMP-2 [5]
57
  • Pro-MMP-3 (only high levels detected >100 ng) [4]
55
  • Pro-MMP-1 (only very high levels detected, like 500 ng) [4]
45
  • MMP-2 after long incubation with APMA [2]
  • Catalytic domain of MMP-2 [4]
  • Mature MMP-3 after incubation with APMA (only high levels detected >100 ng) [4]
43
  • Active MMP-1 (only very high levels detected, like 500 ng) [4]
28
  • Catalytic domain of MMP-3 after incubation with APMA (only high levels detected >100 ng) [4]

References

1.         Snoek-van Beurden, P.A. and J.W. Von den Hoff, Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors. Biotechniques, 2005. 38(1): p. 73-83.

2.         Toth, M. and R. Fridman, Assessment of Gelatinases (MMP-2 and MMP-9 by Gelatin Zymography. Methods in molecular medicine, 2001. 57: p. 163-74.

3.         Koo, B.H., et al., Dimerization of matrix metalloproteinase-2 (MMP-2): functional implication in MMP-2 activation. J Biol Chem, 2012. 287(27): p. 22643-53.

4.         Troeberg, L. and H. Nagase, Measurement of matrix metalloproteinase activities in the medium of cultured synoviocytes using zymography. Methods Mol Biol, 2003. 225: p. 77-87.

5.         Hussain, A.A., Y. Lee, and J. Marshall, High molecular-weight gelatinase species of human Bruch’s membrane: compositional analyses and age-related changes. Investigative ophthalmology & visual science, 2010. 51(5): p. 2363-71.

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