山东大学学报(理学版) ›› 2017, Vol. 52 ›› Issue (9): 103-110.doi: 10.6040/j.issn.1671-9352.0.2017.155
• • 上一篇
许柏英1,2,张甜甜1
XU Bo-ying1,2, ZHANG Tian-tian1
摘要: 铜绿微囊藻(Microcystisaeruginosa PCC 7806)是我国淡水湖泊生态系统季节性蓝藻水华爆发的最优势藻种,其爆发的机制之一在于气囊,它是一种充满气体的蛋白质性质的亚细胞器结构,可为藻细胞提供浮力,调节藻细胞在水体中的位置以利于生长和后续的生境殖民化。铜绿微囊藻的气囊合成涉及14个气囊蛋白,但大部分气囊蛋白在气囊合成过程中的具体分子功能尚不清楚。对其中一个气囊蛋白GvpW进行了分子克隆,利用原核表达系统在体外进行了异源表达,结合镍柱亲和层析和凝胶过滤层析纯化出了高纯度的GvpW,运用动态光散射和化学交联证实在体外纯化后的GvpW是呈单体状态,同时通过晶体初筛和优化获得了生长较好的蛋白质晶体。这为后续解析GvpW的三维结构及在分子水平直观探讨其在气囊合成中的生理功能奠定了基础。
中图分类号:
[1] LARSEN H, OMANG S, STEENSLAND H. On the gas vacuoles of the halobacteria[J]. Archiv Für Mikrobiologie, 1967, 59(1-3): 197-203. [2] PFENNIG N,COHEN-BAZIRE G. Some properties of the green bacterium Pelodictyon clathratiforme[J]. Archiv Für Mikrobiologie, 1967, 59(1): 226-236. [3] STOECKENIUS W, KUNAU W H. Further characterization of particulate fractions from lysed cell envelopes of Halobacterium halobium and isolation of gas vacuole membranes[J]. Journal of Cell Biology, 1968, 38(2): 336-357. [4] WALSBY A E. Gas vesicles[J]. Microbiological Reviews, 1994, 58(1): 94-144. [5] WALSBY A E. Structure and function of gas vacuoles[J]. Bacteriological Reviews, 1972, 36(1): 1-32. [6] WALSBY A E, BUCKLAND B. Isolation and purification of intact gas vesicles from a blue-green alga [J].Nature(London), 1969, 224(1): 716-717. [7] JONES D D, JOST M. Characterization of the protein from gas-vacuole membranes of the blue-green alga, Microcystis aeruginosa[J]. Planta, 1971, 100(4): 277-287. [8] BOWEN C C, JENSEN T E. Blue-green algae: fine structure of the gas vacuoles [J]. Science, 1965, 147(3664): 1460-1462. [9] JOST M. Die Ultrastruktur von Oscillatoria rubescens D. C [J]. Archives of Microbiology, 1965, 50(3): 211-245. [10] WALSBY A E. The permeability of blue-green algal gas-vacuole membranes to gas[J]. Proceedings of the Royal Society B Biological Sciences, 1969, 173(1031):235-255. [11] WALSBY A E. Permeability of gas vesicles to perfluorocyclobutane [J]. Microbiology, 1982, 128(8): 1679-1684. [12] HAYES P K, WALSBY A E. The inverse correlation between width and strength of gas vesicles in cyanobacteria[J]. Brit Phycol J Trevor, 1986, 21(2):191-197. [13] DUNTON P G, WALSBY A E. The diameter and critical collapse pressure of gas vesicles in Microcystis are correlated with GvpCs of different length[J]. FEMS Microbiology Letters, 2005, 247(1): 37-43. [14] PFEIFER F. Distribution, formation and regulation of gas vesicles [J]. Nature Reviews Microbiology, 2012, 10(10): 705-715. [15] MLOUKA A, COMTE K, CASTETS A M, et al. The gas vesicle gene cluster from Microcystis aeruginosa and DNA rearrangements that lead to loss of cell buoyancy [J]. Journal of Bacteriology, 2004, 186(8): 2355-2365. [16] HAYES P K, WALSBY A E, WALKER J E. Complete amino acid sequence of cyanobacterial gas-vesicle protein indicates a 70-residue molecule that corresponds in size to the crystallographic unit cell [J]. Biochemical Journal, 1986, 236(1): 31-36. [17] BUCHHOLZ B E, HAYES P K, WALSBY A E. The distribution of the outer gas vesicle protein, GvpC, on the Anabaena gas vesicle,and its ratio to GvpA [J].Journal of General Microbiology, 1993, 139(10): 2353-2363. [18] HAYES P K, LAZARUS C M, BEES A, et al. The protein encoded by gvpC is a minor component of gas vesicles isolated from the cyanobacteria Anabaena flos-aquae and Microcystis sp[J]. Molecular Microbiology,1988, 2(5): 545-552. [19] WALSBY A E, HAYES P K. The minor cyanobacterial gas vesicle protein, GvpC, is attached to the outer surface of the gas vesicle [J]. Journal of General Microbiology, 1988, 134(10): 2647-2657. [20] ENGLERT C,PFEIFER F. Analysis of gas vesicle gene expression in Haloferax mediterranei reveals that GvpA and GvpC are both gas vesicle structural proteins [J]. Journal of Biological Chemistry, 1993, 268(13): 9329-9336. [21] DUNTON P G, MAWBY W J, SHAW V A, et al. Analysis of tryptic digests indicates regions of GvpC that bind to gas vesicles of Anabaena flos-aquae[J]. Microbiology, 2006, 152(6): 1661-1669. [22] XU Boying, DAI Yanan, ZHOU Kang, et al. Structure of the gas vesicle protein GvpF from the cyanobacterium microcystis aeruginosa[J]. Acta crystallographica. Section D,Biological crystallography, 2014, 70(11):3013-3022 |
[1] | 巨颖琳,李小明*. 南四湖3种沉水植物对铜绿微囊藻化感作用研究[J]. J4, 2011, 46(3): 1-8. |
|