JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2017, Vol. 52 ›› Issue (9): 103-110.doi: 10.6040/j.issn.1671-9352.0.2017.155

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Heterelogous expression, purification andcrystal growth of gas vesicle protein GvpW from the cyanobacterial Microcystis aeruginosa

XU Bo-ying1,2, ZHANG Tian-tian1   

  1. 1. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China;
    2. School of Life Sciences, University of Science and Technology of China, Heifei 230026, Anhui, China
  • Received:2017-04-12 Online:2017-09-20 Published:2017-09-15

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Abstract: Microcystis aeruginosa PCC 7806 is the most dominant algae of seasonal outbreak cyanobacteria bloom in fresh water lake ecosystem of China. One of the outbreak mechanisms for the occurrence of cyanobacterial water-bloom is gas vesicles, which are gas-filled proteinaceous organelles and provide varying buoyancy for cyanobacteria to regulate the position for growth and subsequent colonization. The gas vesicle of M. aeruginosa is composed of 14 gas vesicles proteins, but it is still unclear what the specific molecular functions of most Gvp proteins are during the process of gas vesicle synthesis. In this study, a gvpW gene was cloned and heterologous expressed in Escherichia coli, and the recombinant protein was purified by Ni2+ affinity chromatography and gel filtration. Dynamic light scattering and chemical crosslinking analyses showed that the recombinant protein GvpW mainly existed as monomer in vitro. In addition, GvpW protein crystals were obtained by crystal screening and optimization. These results provide a basis for further studies on its three-dimensional structure determination and molecular functions.

Key words: Microcystis aeruginosa, Gvp proteins, GvpW, protein crystals, gas vesicles

CLC Number: 

  • Q816
[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
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