• 생명과학회지
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• 제23권8호
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• pp.978-983
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• 2013

Kinesin-II는 다양한 운반체들을 미세소관을 따라 운반하는 motor 단백질의 하나이다. Kinesin-II는 두 개의 motor 단백질 KIF3A와 KIF3B, 그리고 motor 단백질의 말단에 결합하는 kinesin superfamily-associated protein 3 (KAP3)로 구성되어 있다. KAP3는 Kinesin-II의 기능에 중요한 역할을 하는 것으로 알려져 있으나 명확한 기능은 아직 밝혀지지 않았다. 본 연구에서 KAP3와 결합하는 단백질을 분리하기 위하여 효모 two-hybrid system을 사용하여 탐색한 결과 HS-1-associated protein X-1 (HAX-1)을 분리하였다. KAP3은 HAX-1의 C-말단 부위와 결합하며, HAX-1은 KAP3의 C-말단부위와 결합함을 효모 two-hybrid assay로 확인하였다. 그러나, HAX-1는 KIF3A, KIF3B, KIF5B, 그리고 kinesin light chain (KLC)과는 결합하지 않았다. KAP3와 HAX-1의 단백질 결합은 glutathione S-transferase (GST) pull-down assay와 공동면역침강으로 추가 확인하였다. 생쥐의 뇌 파쇄액을 HAX-1 항체와 KIF3A 항체로 면역침강을 행한 결과 Kinesin-II의 구성단백질인 KIF3B와 KAP3가 같이 침강하였다. 이러한 결과들은 KAP3가 Kinesin-II와 HAX-1의 결합을 매개한다는 것을 시사한다.

• Journal of Applied Biological Chemistry
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• 제50권2호
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• pp.58-62
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• 2007

Identification of Dual-specificity protein phosphatase (DSP) substrates is essential in revealing physiological roles of DSPs. We isolated VHZ-interacting proteins from extracts of 293T cells overexpressing a VHZ (C95S, D65A) mutant known to be substrate- trapping mutant. Analysis of specific proteins bound to VHZ by 2D gel electrophoresis and mass spectroscopy revealed that these proteins contained Chaperonin containing TCP1, Type II phosphatidylinositol phosphate kinase ${\gamma}$, Intraflagellar transport 80 homolog, and Kinesin superfamily protein 1B. VHZ-interacting proteins showed that VHZ is involved in many important cellular signal pathways such as protein folding, molecular transportation, and tumor suppression.

• 한국산업보건학회지
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• 제20권1호
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• pp.29-40
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• 2010

This study was aimed at investigating the gene expression profile in basal ganglia of cadmium exposed rat based on cDNA array analysis. For cDNA array analysis, adult Sprague-Dawley male rats (350 ${\pm}$ 25 g) were intraperitoneally injected with 2.0 mg/kg body weight/day of CdCl2 (0.3 ml) for 5 days. For doserelated gene expression analysis rats were intraperitoneally injected with 0.0, 0.1, 0.3, 1.0 mg/kg body weight/day of CdCl$_2$ for 5 days. Control rats were injected with equal volume of saline. Cadmium concentration of brain was analyzed by atomic absorption spectrophotometer. For cDNA array, RNA samples were extracted from basal ganglia and reverse-transcribed in the presence of [${\alpha}$32P]-dATP. Membrane sets of the Atlas Rat 1.2 array II and Toxicology array 1.2 (Clontech, Palo Alto, CA) were hybridized with cDNA probe sets. RT-PCR was employed to validate the relative gene expression patterns obtained from the cDNA array. Northern blot hybridization methods were employed to assess the dose-related gene expression. Among the 2352 cDNAs, 671 genes were detected in both array sets and 63 genes of 38 classes showed significant (more than two fold) changes in expression. Thirty five of these genes were up-regulated and twenty eight were down-regulated in the cadmium exposed group. According to the dose-related gene expression analysis, heat shock 27 kDa protein (HSP27), neurodegeneration-associated protein 1 (Neurodap 1) genes were significantly up-regulated and melatonin receptor 1a (Mel1a), Kinesin family member 3C (KIF3C), novel kinesinrelated protein (KIF1D) genes were significantly downregulated even in the low-dose of cadmium exposed group (0.1 mg/kg body weight/day). Conclusions Sixty three genes detected in this study can give some more useful informations about the cadmium-induced neurotoxicity in the basal ganglia. As well as, HSP27, Neurodap1, Mel1a, KIF3C and KIF1D genes may be useful for the study of the cadmium-induced neurotoxicity because these genes showed dramatic changes of mRNA levels in response to the low dose of cadmium exposure.