Shirai Y, Uno T, Aizono Y
Laboratory of Molecular Pharmacology, Kobe University, Hyogo, Japan.
Arch Insect Biochem Physiol. 1998;38(4):177-84. doi: 10.1002/(SICI)1520-6327(1998)38:4<177::AID-ARCH3>3.0.CO;2-O.
At least three GTP-binding proteins (G-proteins), 28, 25, and 21 kDa, were found in the brain-corpus cardiacum-corpus allatum complex (BR-CC-CA) of the silkworm, Bombyx mori. They bound to GTP and GDP specifically among nucleotides tested, indicating that these proteins are small G-proteins. The 25 kDa G-protein showed a cross-reactivity to anti-rab3A antibody, while it did not cross-react with anti-rhoA, rab3B, and anti-ras antibodies. On the other hand, the 28 and 21 kDa G-proteins showed no cross-reactivity to any of those antibodies tested. Immunoblot analysis using the anti-rab3A antibody demonstrated that the 25 kDa G-protein was detected preferentially in the BR-CC-CA, and to some extent in the suboesophageal ganglion, but not in the salivary gland, fat body, prothoracic gland, and oesophagus. These results suggested that the 25 kDa G-protein was a member of the rab family of G-proteins. Furthermore, 1 mM GTP gamma S capable of activating G-proteins induced BR-CC-CA to release PTTH under the conditions that stimulation of the PTTH release with hetero-trimeric G-protein was suppressed. These results indicated that the small G-proteins may possibly contribute to PTTH release in Bombyx mori.
在家蚕(Bombyx mori)的脑-心侧体-咽侧体复合体(BR-CC-CA)中发现了至少三种GTP结合蛋白(G蛋白),分子量分别为28 kDa、25 kDa和21 kDa。在所测试的核苷酸中,它们特异性地与GTP和GDP结合,表明这些蛋白是小G蛋白。25 kDa的G蛋白与抗rab3A抗体有交叉反应,而与抗rhoA、rab3B和抗ras抗体无交叉反应。另一方面,28 kDa和21 kDa的G蛋白与所测试的任何一种抗体均无交叉反应。使用抗rab3A抗体进行的免疫印迹分析表明,25 kDa的G蛋白优先在BR-CC-CA中被检测到,在咽下神经节中也有一定程度的检测,但在唾液腺、脂肪体、前胸腺和食管中未检测到。这些结果表明,25 kDa的G蛋白是G蛋白rab家族的成员。此外,在异源三聚体G蛋白刺激PTTH释放受到抑制的条件下,能够激活G蛋白的1 mM GTPγS诱导BR-CC-CA释放PTTH。这些结果表明,小G蛋白可能在家蚕PTTH释放中发挥作用。