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Using substrate‐binding variants of the cAMP‐dependent protein kinase (PKA) to identify docking sites within two physiological target proteins
Author(s) -
Deminoff Stephen J.,
Ramachandran Vidhya,
Herman Paul K.
Publication year - 2008
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.22.2_supplement.321
Subject(s) - phosphorylation , docking (animal) , signal transduction , kinase , protein kinase a , microbiology and biotechnology , biology , computational biology , chemistry , biochemistry , medicine , nursing
Cells use complex signaling networks to bring about the appropriate responses to stimuli and defects in these signaling pathways contribute to cancer and other human diseases. A full understanding of signal transduction mechanisms requires insight into the ways in which protein kinases mediate this signaling activity. The specificity and efficiency with which physiological substrates are phosphorylated involves regions of substrates that are distal to the site(s) of phosphorylation. Our lab has identified catalytically impaired variants of PKA that bind stably and specifically to substrates. We have used these variants to identify both candidate docking sites within substrates, and the subdomains of PKA with which they interact. Importantly, disrupting these putative docking interactions impairs phosphorylation both in vitro and in vivo . Precisely defining these kinase‐substrate interactions will help to elucidate signaling pathways in general and may also identify novel points for therapeutic intervention. This work is supported by a grant from the National Institutes of Health (GM65227).