Technical Brief: a novel strategy for enrichment of trabecular meshwork protease proteome.

ABSTRACT: We present a novel and simple enrichment strategy to capture trabecular meshwork (TM) protease proteome. The method relies on fractionation of TM tissue into cytosolic and nuclear extracts and subsequent affinity enrichment of proteases on peptide inhibitors. A large repertoire of available protease substrate analog peptides enables an improved capture of TM protease proteome compared to SDS-PAGE fractionation alone. Peptide analog inhibitors of protease substrates are immobilized on a protein A or G column using 254 nm intense ultraviolet (UV) light. The TM cytosolic protein extract incubated on the column is eluted with salt or a buffer with a low hydrogen ion concentration. The resultant protein solution is precipitated with acetone, fractionated on SDS-PAGE, in situ trypsin digested, a

SUBMITTER: Picciani R 

PROVIDER: S-EPMC2386506 | biostudies-literature | 2008 May

REPOSITORIES: biostudies-literature