Peng Xiaozhong

Cloning and prokaryotic expression of HGLP cDNA

A novel cDNA,HGLP, which encodes a G-protein coupled receptor (GPCR) like protein, has been isolated and cloned. The coding region of the humanHGLP predicts a 7-transmembrane region protein with motifs of rhodopsin-like GPCR superfamily. Northern blot analysis reveals a 3-kb transcript in various human tissues examined. The N- and C-terminal coding regions ofHGLP, which are deduced as non-transmembrane regions, have been amplified by PCR and cloned into pET30a+ vector. Then the recombinant proteins are highly expressed inE. coli.

Human nectin—like 1, a novel membrane protein interacting with protein 4.1 R

Human nectin—like 1 (NECL1) full—length cDNA was cloned by bioinformatics method when searching for candidate membrane proteins interacting with members of protein 4.1 family. The cytoplasmic and extracellular regions of NECLl were expressed in and purified from E.coli, and the polyclonal antibody was produced. Interaction between the cytoplasmic region of NECLl and the 30 kD membrane binding domain of protein 4.1 on red blood cell (4.1R) was demonstrated by IAsys—biosensor system and GST pull—down experiment. Results of biotin—labeled peptide ELISA further demonstrated the key amino acids for the binding. The interaction research of NECLls cytoplasmic domain provides basis for further study of the functions of NECL1 in nervous system. Supported by National Natural Science Foundation of China (Grant No. 50271027), Fujian International Cooperation Foundation (Grant No.20021018), and Fujian Natural Science Foundation (Grant Nos. E0310023 and 2002J010)