RT Journal
A1 Kaberdin, Vladimir R.
A1 McDowall, Kenneth J.
T1 Expanding the Use of Zymography by the Chemical Linkage of Small, Defined Substrates to the Gel Matrix
JF Genome Research 
JO Genome Research 
YR 2003 
FD August 01 
VO 13 
IS 8 
SP 1961 
OP 1965 
DO 10.1101/gr.1277303 
UL http://genome.cshlp.org/content/13/8/1961.abstract 
AB In the postgenomic era, the comprehensive proteomic analysis of metabolic  and signaling pathways is inevitably faced with the challenge of large-scale  identification and characterization of polypeptides with a particular  enzymatic activity. Previous work has shown that a wide variety of enzymatic  activities of microbial, plant, and animal origin can be assigned to  individual polypeptides using in-gel activity staining (zymography). However,  a number of limitations, such as special substrate requirements, the lack of a  standard procedure, and difficulties in distinguishing enzymes with  overlapping activities have precluded the widespread use of zymography as a  routine laboratory method. Here we demonstrate that, by employing  small-defined substrates that are covalently attached to the gel matrix, we  can largely overcome the aforementioned problems and assay readily a number of  different classes of enzymatic activities within gels after standard  SDS-polyacrylamide electrophoresis. Moreover, this development is compatible  with the two-dimensional separation of proteins and thus has great potential  in the high-throughput screening and characterization of complex biological  and clinical samples.