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Purification and identification of Polyisoprenylated Methylated Protein Methyl Esterase from Porcine Brain
Author(s) -
Duverna Randolph,
Aguilar Byron J,
Amissah Felix,
Lamango Nazarius S
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.776.1
Subject(s) - chemistry , esterase , gene isoform , biochemistry , enzyme , trypsin , isozyme , endoplasmic reticulum , western blot , microbiology and biotechnology , biology , gene
Polyisoprenylated methylated protein methyl esterase (PMPMEase) is a key enzyme in the polyisoprenylation pathway (PP). This PP is important for the post‐translational modification of proteins which regulate cell growth, differentiation and apoptosis. Porcine Liver Esterase (PLE), an isoform of PMPMEase has been isolated and characterized. A Recent study strongly suggests the existence of a brain isoform of PMPMEase. The principal objective was to isolate, identify and characterize the brain isoform of porcine liver esterase. Using various chromatographic techniques, brain PMPMEase was partially purified to few bands. Active fractions were analyzed by SDS‐PAGE and the coomassie bands digested with trypsin, and analyzed using MALDI‐TOF/TOF Mass Spectrometry (MS) followed by Mascot database searches. The enzyme was identified from SwissProt, NCBI and MSDB databases as an endoplasmic reticulum, single‐pass calcium‐independent phospholipase A2. This is interesting given the similar characteristics between the known lipid substrates of the enzyme and the polyisoprenyl moieties of the putative protein substrates. This information will now be used to produce recombinant enzyme for substrate and inhibitors kinetics studies.