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Transfection of cDNA with G→T point mutation at the cleavage site of insulin receptors to cos 7 cells

Kobayashi, Masashi ; Sugibayashi, Masaaki ; Sasaoka, Toshiyasu ; Egawa, Katsuya ; Shigeta, Yuikio ; Tamaki, Mikio ; Nakamura, Etsuo ; Teraoka, Hiroshi

Biochemical and biophysical research communications, 1990-03, Vol.167 (3), p.1073-1078 [Periódico revisado por pares]

United States: Elsevier Inc

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Título:
Transfection of cDNA with G→T point mutation at the cleavage site of insulin receptors to cos 7 cells
Autor: Kobayashi, Masashi ; Sugibayashi, Masaaki ; Sasaoka, Toshiyasu ; Egawa, Katsuya ; Shigeta, Yuikio ; Tamaki, Mikio ; Nakamura, Etsuo ; Teraoka, Hiroshi
Assuntos: Amino Acid Sequence ; Animals ; Cell Line ; DNA - genetics ; Gene Expression ; Genetic Vectors ; Humans ; Insulin Resistance - genetics ; Kinetics ; Molecular Sequence Data ; Mutation ; Plasmids ; Receptor, Insulin - biosynthesis ; Receptor, Insulin - genetics ; Receptor, Insulin - metabolism ; Transfection
É parte de: Biochemical and biophysical research communications, 1990-03, Vol.167 (3), p.1073-1078
Descrição: To study whether the G→T point mutation of insulin proreceptors at the cleavage site which changed -Arg-Lys-Arg-Arg- to -Arg-Lys-Arg-Ser- caused unprocessed insulin receptors with decreased insulin binding affinity, we performed transfection of cDNA with the mutation in COS 7 cells and examined the expressed insulin receptors. After site-directed mutagenesis, an expression vector pGEM3SV was used to make a plasmid which contained full-length HIRcDNA behind SV40 early promoter. Transfection of normal HIR cDNA produced normal insulin receptors on the plasma membranes in COS 7 cells. However, transfection of cDNA with the mutation resulted in the presence of 210K proreceptors in the plasma membranes with decreased insulin binding ability (35% of normal). These results suggest that the mutation, not the defect of converting enzyme, was the cause for unprocessed insulin proreceptors in the patients with insulin resistance.
Editor: United States: Elsevier Inc
Idioma: Inglês

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Transfection of cDNA with G→T point mutation at the cleavage site of insulin receptors to cos 7 cells

Kobayashi, Masashi ; Sugibayashi, Masaaki ; Sasaoka, Toshiyasu ; Egawa, Katsuya ; Shigeta, Yuikio ; Tamaki, Mikio ; Nakamura, Etsuo ; Teraoka, Hiroshi

United States: Elsevier Inc

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