Expression and characterization of kinase-active v-erbB protein using a baculovirus vector system.
The v-erbB gene is an oncogene of the avian erythroblastosis virus encoding a protein that is a truncated version of the epidermal growth factor receptor. The v-erbB protein was expressed alone or as polyhedrin-erbB fusion proteins using the Bombyx mori nuclear polyhedrosis virus vector. The expression level of the fusion protein whose polyhedrin portion consisted of only 8 amino-terminal amino acids was more than ten times higher than that of the non-fusion protein. Studies with tunicamycin showed that the recombinant v-erbB proteins were glycosylated. The recombinant protein autophosphorylated tyrosine residues, and phosphorylated a synthetic tyrosine-containing peptide and lipocortin I. These observations indicate that functional v-erbB protein can be expressed in silkworm-derived cells, and furthermore, that this system can be used for large-scale production.
Exploratory Research Laboratories 2, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
K
K
Morishita
M
M
Iwamoto
K
K
Murakami
M
M
Kubota
S
S
Maeda
K
K
Toyoshima
T
T
Yamamoto
D000595:Amino Acid Sequence; D017302:Annexins; D016367:Baculoviridae / Q000235:genetics*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D005822:Genetic Vectors; D006031:Glycosylation; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D018739:Oncogene Proteins v-erbB; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D015682:Retroviridae Proteins, Oncogenic / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D017302:Annexins; D002135:Calcium-Binding Proteins; D009842:Oligopeptides; D018739:Oncogene Proteins v-erbB; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D015682:Retroviridae Proteins, Oncogenic; D011505:Protein-Tyrosine Kinases