Science 重組膜蛋白 生物化學(xué)系與分子藥理學(xué)系的科學(xué)家近期在膜蛋白研究方面取得新進(jìn)展,β-Barrel膜蛋白位于革蘭氏陰性細(xì)菌、線粒體和葉綠體中,這種膜蛋白由高度保守和復(fù)雜的多蛋白復(fù)合體組成。β-Barrel膜蛋白的分子結(jié)構(gòu)、折疊結(jié)構(gòu)以及與酶作用的部位的結(jié)構(gòu)機(jī)制卻鮮少為人所知。研究β-Barrel膜蛋白的瓶頸在于,β-Barrel膜蛋白很難在一個(gè)生物化學(xué)體系重重新構(gòu)建起來。 β-Barrel膜蛋白的折疊是在一個(gè)沒有能量的環(huán)境下進(jìn)行的,其中一種可溶性的伴侶分子是β-Barrel膜蛋白折疊的關(guān)鍵因素。 膜蛋白 膜蛋白是指能夠結(jié)合或整合到細(xì)胞或細(xì)胞器的膜上的蛋白質(zhì)的總稱。而細(xì)胞中一半以上的蛋白質(zhì)可以與膜以不同形式結(jié)合。根據(jù)與膜結(jié)合強(qiáng)度的不同,膜蛋白可以被分為兩類:外周膜蛋白和內(nèi)在膜蛋白。 內(nèi)在膜蛋白是整合于膜上的蛋白質(zhì),總是與膜結(jié)合在一起??梢远x為需要通過人工加入去垢劑(如SDS或Triton X-100)或其他非極性溶劑才能夠從膜中分離出來的蛋白質(zhì)。內(nèi)在膜蛋白還可以根據(jù)與雙分子膜之間結(jié)合關(guān)系的差異細(xì)分為: 跨膜蛋白,顧名思義即跨越膜的兩端的蛋白質(zhì),其跨膜部分為β桶或α螺旋結(jié)構(gòu) 單向內(nèi)在膜蛋白,其只從一個(gè)方向(膜外或膜內(nèi))與膜結(jié)合,雖然部分插入膜中,但不跨膜。 Reconstitution of Outer Membrane Protein Assembly from Purified Components Christine L. Hagan,1,* Seokhee Kim,1,*, Daniel Kahne1,2, β-Barrel membrane proteins in Gram-negative bacteria, mitochondria, and chloroplasts are assembled by highly conserved multiprotein complexes. The mechanism by which these molecular machines fold and insert their substrates is poorly understood. It has not been possible to dissect the folding and insertion pathway because the process has not been reproduced in a biochemical system. We purified the components that fold and insert E. coli outer membrane proteins and reconstituted β-barrel protein assembly in proteoliposomes using the enzymatic activity of a protein substrate to report on its folding state. The assembly of this protein occurred without an energy source, but required a soluble chaperone in addition to the multiprotein assembly complex. |