Diversity of G proteins in Lepidopteran cell lines: partial sequences of six G protein alpha subunits.

Abstract

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.