Identification of protein kinase C inhibitory activity associated with a polypeptide isolated from a phage display system with homology to PCM-1, the pericentriolar material-1 protein

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DOIResolve DOI: http://doi.org/10.1016/j.bbrc.2012.06.093
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TypeArticle
Journal titleBiochemical and Biophysical Research Communications
ISSN0006-291X
Volume424
Issue1
Pages147151; # of pages: 5
SubjectProtein kinase C (PKC); Inhibitor; Phage display; Pericentriolar material-1 (PCM-1) protein
AbstractWe had previously identified a protein kinase C (PKC) inhibitor in murine neuroblastoma cells (Chakravarthy et al. [1]). Similar PKC inhibitory activity was also found in adult rat brain. Using polyclonal antibodies raised against the partially purified PKC inhibitor from rat brain as bait, we isolated a putative brain PKC inhibitor using a T-7 phage display system expressing human brain cDNA library. After enriching the phage population expressing the putative PKC inhibitor with four rounds of biopanning using ELISA and in vitro PKC binding assays, we identified a phage clone that expressed a product with significant PKC inhibitory activity. We have cloned and expressed this cDNA in a bacterial system and purified the recombinant protein. This polypeptide (174 amino acids) is highly homologous to a region of the 228-kDa PCM-1, the human pericentriolar material 1 protein. We have mapped this polypeptide’s PKC-inhibitory domain and shown its PKC inhibitory activity in vitro. However, it will need to be determined whether the full-length PCM-1 protein possesses PKC inhibitory activity in vivo, and if so, how this might contribute to PCM-1’s recently demonstrated roles in ciliogenesis and neurogenesis.
Publication date
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
Peer reviewedYes
IdentifierS0006291X12011886
NPARC number21268673
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Record identifierbc88feb5-e45f-475d-8452-7c25e140a162
Record created2013-11-07
Record modified2016-05-09
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