Clinical Pharmacology & Biopharmaceutics
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GPCRs are large family of cell surface proteins participating in signal transduction where C-terminus recruits GPCR Interacting
Proteins (GIPs) that regulate GPCR function. The 5-HT
(a, d, e, f and g) receptor (R) splice variants possess canonical type 1
or type 2 PDZ domains predicted to interact with various GIPs which could influence their modulatory and prokinetic functions
in the intestine.
This study investigates the distribution of 5-HT
R and GIPs in the guinea pig intestine and to determine if human 5-HT
and specific GIPs interact using
cell system. RT-PCR, western and immunoflourescence analysis were used to investigate
transcript and protein expression. N-terminal FLAG tagged 5-HT
R variants and Lin 7 homologues (h) with C-terminal tagged
V5, c-Myc and HA constructs were generated and expressed in COS-7 cells to study interactions.
R, GRKs and Lin 7h were expressed in the guinea pig intestine. Lin 7h and 5-HT
R co-localized in cell lines and the
aR and Lin 7h were co-immunoprecipitated.
We have previously shown that 5-HT
R and GIPs, GRKs and Lin7h, are found in the human colon. Our data indicates that
this occurs in the guinea pig and we demonstrate that it may be possible for 5-HT
R and Lin 7h to interact in the same cells. The
results suggest Lin 7h could be a potential target to modulate 5-HT
R function. This would be of particular relevance when 5-HT
variant expression is altered as demonstrated in cancerous tissue (Cartier et al. 2005).
Kenneth Chinkwo completed his Ph.D. degree in Molecular Pharmacology at Monash University Australia in 2011. He studied serotonin receptors and G-protein coupled receptor (GPCR) interacting proteins; their role as potential therapeutic targets in clinical medicine. Ken holds a B.Sc. hons in Medical Laboratory Sciences at the University of Calabar Teaching hospital (UCTH) Nigeria. He worked in several pathology laboratories in Cameroon his home country. Ken obtained a Masters Degree in Biochemistry in the area of cancer biology in the University of the Western Cape, South Africa. Currently, he is a Medical Science lecturer in Charles Sturt University (CSU) Australia.
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