Diversity of G proteins in Lepidopteran cell lines: partial sequences of six G protein alpha subunits.
The aim of this work was to sample the diversity of G protein alpha subunits in lepidopteran insect cell lines. Here we report the amplification by degenerate PCR of partial sequences representing six G protein alpha subunits from three different lepidopteran insect cell lines. Sequence comparisons with known G protein alpha subunits indicate that the Sf9, Ld and High Five cell lines each contain (at least) one Galpha(q)-like and one Galpha(i)-like Galpha subunit. All six PCR products are unique at the nucleotide level, but the translation products of the three Galpha q-like partial clones (Sf9-Galpha 1, Ld-Galpha 1, and Hi5-Galpha 1) are identical, as are the translation products of the three Galpha i-like partial clones (Sf9-Galpha 2, Ld-Galpha 2, and Hi5-Galpha 2). Both the Galpha(q)-like and Galpha(i)-like translation products are identical to known Galpha subunits from other Lepidoptera, are highly similar (88-98%) to Galpha subunits from other invertebrates including mosquitoes, fruit flies, lobsters, crabs, and snails, and are also highly similar (88-90%) to known mammalian Galpha subunits. Identification of G protein alpha subunits in lepidopteran cell lines will assist in host cell line selection when insect cell lines are used for the pharmacological analysis of human GPCRs.
Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.
Peter J K
PJ
Knight
Thomas A
TA
Grigliatti
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics* / Q000378:metabolism; D014644:Genetic Variation; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018076:DNA, Complementary; D044385:GTP-Binding Protein alpha Subunits