Small GTP binding proteins: Rab GTPases from the brain of Bombyx mori
Copyright policy )Small GTP binding proteins: Rab GTPases from the brain of Bombyx mori
AbstractFrom a mRNA of the brain of Bombyx mori, we isolated 8 cDNA clones (BRabs), each of which encodes a different member of Rab‐protein family. Four of them have more than 80% amino acid identity to the corresponding members of Drosophila Rab proteins. The other 4 proteins show low sequence similarity to any of the known Rab proteins. However, all of them contain the region conserved in rab protein. Using RACE (Rapid Amplification of cDNA ends), the one full‐length cDNA clone (BRab14) was isolated. The clone was expressed in Escherichia coli as a glutathione S‐transferase (GST) fusion protein. After purification, the fusion protein was cut with protease to remove GST‐Tag and applied to a glutathione S‐Sepharose column. The protein bound [3H]‐GDP with association constant of 1.02 × 1011 M–1. Further, the protein was phosphorylated by protein kinase. This result suggests that Rab protein in the brain of Bombyx mori binds GDP or GTP and its function is regulated by phosphorylation. Arch. Insect Biochem. Physiol. 52:130–138, 2003. © 2003 Wiley‐Liss, Inc.
- Kobe University Japan
DNA, Complementary, Base Sequence, Reverse Transcriptase Polymerase Chain Reaction, Molecular Sequence Data, Brain, Bombyx, Guanosine Diphosphate, Recombinant Proteins, rab GTP-Binding Proteins, Escherichia coli, Animals, Insect Proteins, Amino Acid Sequence, Cloning, Molecular, Phosphorylation, Sequence Alignment, Protein Binding
DNA, Complementary, Base Sequence, Reverse Transcriptase Polymerase Chain Reaction, Molecular Sequence Data, Brain, Bombyx, Guanosine Diphosphate, Recombinant Proteins, rab GTP-Binding Proteins, Escherichia coli, Animals, Insect Proteins, Amino Acid Sequence, Cloning, Molecular, Phosphorylation, Sequence Alignment, Protein Binding
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