生物化学与生物物理进展
生物化學與生物物理進展
생물화학여생물물리진전
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2008年
6期
719-723
,共5页
王健%朱嗣博%李鼎%朱乃硕
王健%硃嗣博%李鼎%硃迺碩
왕건%주사박%리정%주내석
Rab26%RabGTP酶%内吞作用
Rab26%RabGTP酶%內吞作用
Rab26%RabGTP매%내탄작용
Rab26%Rab GTPases%endocytosis
Rab GTPase家族成员基因及其调节囊泡膜运输途径功能已有许多报道,但Rab26蛋白分子的结构和功能仍不清晰.通过生物信息学分析,并在人脑cDNA文库中克隆了一个新的Rab26基因全长cDNA序列,长1 656bp,与已发表的基因序列相比,在48～50位插入了GCC,在956位T被C取代,而在1 197位G被A取代.该序列包含一个771 bp完整的开放阅读框(ORE),编码256个氨基酸残基的Rab26蛋白,分子质量为27.9 ku(GenBank登录号No.AY646153),而非如以往报告的190个氨基酸残基.GFP荧光标记全长Rb26在哺乳动物细胞中表达显示,Rab26主要呈现在细胞膜状结构相联系的分布,发现该基因高表达能显著增强PE标记的红色异源蛋白质的吞噬.还应用逆转录.聚合酶链反应对多种人肿瘤细胞Rb26表达进行了研究,结果显示,Rab26在Acc2、SPC-A1,K562以及HeLa等肿瘤细胞株呈高表达,而在SMMOL/LC-7721、HepG2、Caco2等肝和肠上皮细胞株中则不表达,值得进一步深入研究.
Rab GTPase傢族成員基因及其調節囊泡膜運輸途徑功能已有許多報道,但Rab26蛋白分子的結構和功能仍不清晰.通過生物信息學分析,併在人腦cDNA文庫中剋隆瞭一箇新的Rab26基因全長cDNA序列,長1 656bp,與已髮錶的基因序列相比,在48～50位插入瞭GCC,在956位T被C取代,而在1 197位G被A取代.該序列包含一箇771 bp完整的開放閱讀框(ORE),編碼256箇氨基痠殘基的Rab26蛋白,分子質量為27.9 ku(GenBank登錄號No.AY646153),而非如以往報告的190箇氨基痠殘基.GFP熒光標記全長Rb26在哺乳動物細胞中錶達顯示,Rab26主要呈現在細胞膜狀結構相聯繫的分佈,髮現該基因高錶達能顯著增彊PE標記的紅色異源蛋白質的吞噬.還應用逆轉錄.聚閤酶鏈反應對多種人腫瘤細胞Rb26錶達進行瞭研究,結果顯示,Rab26在Acc2、SPC-A1,K562以及HeLa等腫瘤細胞株呈高錶達,而在SMMOL/LC-7721、HepG2、Caco2等肝和腸上皮細胞株中則不錶達,值得進一步深入研究.
Rab GTPase가족성원기인급기조절낭포막운수도경공능이유허다보도,단Rab26단백분자적결구화공능잉불청석.통과생물신식학분석,병재인뇌cDNA문고중극륭료일개신적Rab26기인전장cDNA서렬,장1 656bp,여이발표적기인서렬상비,재48～50위삽입료GCC,재956위T피C취대,이재1 197위G피A취대.해서렬포함일개771 bp완정적개방열독광(ORE),편마256개안기산잔기적Rab26단백,분자질량위27.9 ku(GenBank등록호No.AY646153),이비여이왕보고적190개안기산잔기.GFP형광표기전장Rb26재포유동물세포중표체현시,Rab26주요정현재세포막상결구상련계적분포,발현해기인고표체능현저증강PE표기적홍색이원단백질적탄서.환응용역전록.취합매련반응대다충인종류세포Rb26표체진행료연구,결과현시,Rab26재Acc2、SPC-A1,K562이급HeLa등종류세포주정고표체,이재SMMOL/LC-7721、HepG2、Caco2등간화장상피세포주중칙불표체,치득진일보심입연구.
Rab GTPases serve as master regulators of vesicular membrane transport on both the exo-and endocytic pathways. Though there are many reports on Rab proteins, the function of these small proteins still remain in speculation. And no report has ever clarified the character of human Rab26. Here it was reported that a novel Rab protein Rab26 is membranous organelle related and in volved inendocytosis of HeLa cells. By using RT-PCR method a novel Rab26 cDNA full-length cDNA of Rab26 that is 1656 bp was identified.The cDNA sequence that at 1197 is 'A' other than 'G', while 'C' at 956 substitutes for 'T', and has 'GCC' insertion at 48 to 50 compared with published sequences. The complete open reading frame (ORF) is 771 bp in length encoding 256-residue protein with a calculated molecular mass of 27.9 ku (GenBank accession No.AY646153), rather than a shorter one with 190-amino acid residue as reported previously. GFP labeled full-length Rab26 expression showed that Rab26 was mainly sublocated in membranous organelles and could enhance endocytosis which means could took PE labeled protein as an endocytic tracer. RT-PCR analysis showed Rab26 was detected to express in several kinds of adenocarcinoma cell lines such as Acc2, AccM, SPC-A1 and HeLa cell lines, which indicated that Rab26 expression might be associated with some carcinomas.
