Molecular Cloning and Characterization of a Muscle-Specific Lipase from the Bumblebee Bombus ignitus
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A muscle-specific lipase gene of the bumblebee Bombus ignitus was cloned and characterized. This gene, which we named Bi-Lipase, consists of seven exons encoding 317 amino acid residues. Bi-Lipase possesses all the features of lipases, including GXSXG consensus motif and Ser-Asp-His catalytic triad. Expressed as a 37-kDa polypeptide in baculovirus-infected insect Sf9 cells, recombinant Bi-Lipase showed an optimal pH of 9.0 and exhibited its highest catalytic activity at 40oC. Furthermore, through the addition of tunicamycin to the recombinant virus-infected Sf9 cells, recombinant Bi-Lipase was found to be N-glycosylated. Northern and western blot analyses indicated that Bi-Lipase was expressed in the wing, thorax, and leg muscles. These results show that Bi-Lipase is a muscle-specific lipase, suggesting a possible role of Bi-Lipase in the utilization of lipids for muscular activity in B. ignitus.
Abstract
A muscle-specific lipase gene of the bumblebee Bombus ignitus was cloned and characterized. This gene, which we named Bi-Lipase, consists of seven exons encoding 317 amino acid residues. Bi-Lipase possesses all the features of lipases, including GXSXG consensus motif and Ser-Asp-His catalytic triad. Expressed as a 37-kDa polypeptide in baculovirus-infected insect Sf9 cells, recombinant Bi-Lipase showed an optimal pH of 9.0 and exhibited its highest catalytic activity at 40oC. Furthermore, through the addition of tunicamycin to the recombinant virus-infected Sf9 cells, recombinant Bi-Lipase was found to be N-glycosylated. Northern and western blot analyses indicated that Bi-Lipase was expressed in the wing, thorax, and leg muscles. These results show that Bi-Lipase is a muscle-specific lipase, suggesting a possible role of Bi-Lipase in the utilization of lipids for muscular activity in B. ignitus.
참고문헌 (28)
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