Solution and electron microscopy characterization of lactococcal phage baseplates expressed in Escherichia coli

被引：31

作者：

Campanacci, Valerie ^{[1
,2
,3
]}

Veesler, David ^{[1
,2
,3
]}

Lichiere, Julie ^{[1
,2
,3
]}

Blangy, Stephanie ^{[1
,2
,3
]}

Sciara, Giuliano ^{[1
,2
,3
]}

Moineau, Sylvain ^{[4
,5
,6
]}

van Sinderen, Douwe ^{[7
,8
]}

Bron, Patrick ^{[9
]}

Cambillau, Christian ^{[1
,2
,3
]}

机构：

[1] CNRS, UMR 6098, F-13288 Marseille 09, France

[2] Univ Aix Marseille 1, F-13288 Marseille, France

[3] Univ Aix Marseille 2, F-13288 Marseille 09, France

[4] Univ Laval, Fac Med Dent, GREB, Quebec City, PQ G1V 0A6, Canada

[5] Univ Laval, Fac Med Dent, Felix dHerelle Reference Ctr Bacterial Viruses, Quebec City, PQ G1V 0A6, Canada

[6] Univ Laval, Fac Sci & Genie, Dept Biochim & Microbiol, Quebec City, PQ G1V 0A6, Canada

[7] Natl Univ Ireland, Dept Microbiol, Cork, Ireland

[8] Natl Univ Ireland Univ Coll Cork, Alimentary Pharmabiot Ctr, Cork, Ireland

[9] CNRS, INSERM, Ctr Biochim Struct, U554,UMR 5048, F-34090 Montpellier, France

来源：

JOURNAL OF STRUCTURAL BIOLOGY | 2010年 / 172卷 / 01期

基金：

爱尔兰科学基金会; 加拿大自然科学与工程研究理事会;

关键词：

Operon expression; Lactococcus lactis phage; Multi-protein complex; Multi-angle light scattering; Receptor binding protein; Baseplate; Electron microscopy; RECEPTOR-BINDING PROTEIN; SECONDARY STRUCTURE; CIRCULAR-DICHROISM; STRUCTURAL GENOMICS; CRYSTAL-STRUCTURE; DNA-SEQUENCE; BACTERIOPHAGE TP901-1; MODULAR STRUCTURE; GENE-EXPRESSION; LACTIS PHAGES;

D O I：

10.1016/j.jsb.2010.02.007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report here the characterization of several large structural protein complexes forming the baseplates (or part of them) of Siphoviridae phages infecting Lactococcus lactis: TP901-1, Tuc2009 and p2. We revisited a "block cloning" expression strategy and extended this approach to genomic fragments encoding proteins whose interacting partners have not yet been clearly identified. Biophysical characterization of some of these complexes using circular dichroism and size exclusion chromatography, coupled with on-line light scattering and refractometry, demonstrated that the over-produced recombinant proteins interact with each other to form large (up to 1.9 MDa) and stable baseplate assemblies. Some of these complexes were characterized by electron microscopy confirming their structural homogeneity as well as providing a picture of their overall molecular shapes and symmetry. Finally, using these results, we were able to highlight similarities and differences with the well characterized much larger baseplate of the myophage T4. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：75 / 84

页数：10

共 59 条

[1] Implementation of semi-automated cloning and prokaryotic expression screening: the impact of SPINE [J].

Alzari, Pedro M. ;

Berglund, H. ;

Berrow, N. S. ;

Blagova, E. ;

Busso, D. ;

Cambillau, C. ;

Campanacci, V. ;

Christodoulou, E. ;

Eiler, S. ;

Fogg, M. J. ;

Folkers, G. ;

Geerlof, A. ;

Hart, D. ;

Haouz, A. ;

Herman, M. D. ;

Macieira, S. ;

Nordlund, P. ;

Perrakis, A. ;

Quevillon-Cheruel, S. ;

Tarandeau, F. ;

van Tilbeurgh, H. ;

Unger, T. ;

Luna-Vargas, M. P. A. ;

Velarde, M. ;

Willmanns, M. ;

Owens, Raymond J. .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2006, 62 :1103-1113

[2] DETERMINATION OF PROTEIN SECONDARY STRUCTURE IN SOLUTION BY VACUUM ULTRAVIOLET CIRCULAR-DICHROISM [J].

BRAHMS, S ;

BRAHMS, J .

JOURNAL OF MOLECULAR BIOLOGY, 1980, 138 (02) :149-178

[3] Analysis of the complete DNA sequence of the temperate bacteriophage TP901-1: Evolution, structure, and genome organization of lactococcal bacteriophages [J].

Brondsted, L ;

Ostergaard, S ;

Pedersen, M ;

Hammer, K ;

Vogensen, FK .

VIROLOGY, 2001, 283 (01) :93-109

[4] An unusual tryptophanyl tRNA synthetase interacts with nitric oxide synthase in Deinococcus radiodurans [J].

Buddha, MR ;

Keery, KM ;

Crane, BR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (45) :15881-15886

[5] Structural genomics of the SARS coronavirus:: cloning, expression, crystallization and preliminary crystallographic study of the Nsp9 protein [J].

Campanacci, V ;

Egloff, MP ;

Longhi, S ;

Ferron, F ;

Rancurel, C ;

Salomoni, A ;

Durousseau, C ;

Tocque, F ;

Brémond, N ;

Dobbe, JC ;

Snijder, EJ ;

Canard, B ;

Cambillau, C .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2003, 59 :1628-1631

[6] The translation of recombinant proteins in E-coli can be improved by in silico generating and screening random libraries of a 70/96 mRNA region with respect to the translation initiation codon [J].

Care, S. ;

Bignon, C. ;

Pelissier, M. C. ;

Blanc, E. ;

Canard, B. ;

Coutard, B. .

NUCLEIC ACIDS RESEARCH, 2008, 36 (01)

[7] Structure of the outer membrane complex of a type IV secretion system [J].

Chandran, Vidya ;

Fronzes, Remi ;

Duquerroy, Stephane ;

Cronin, Nora ;

Navaza, Jorge ;

Waksman, Gabriel .

NATURE, 2009, 462 (7276) :1011-U66

[8] Analysis of the DNA sequence, gene expression, origin of replication and modular structure of the Lactococcus lactis lytic bacteriophage sk1 [J].

Chandry, PS ;

Moore, SC ;

Boyce, JD ;

Davidson, BE ;

Hillier, AJ .

MOLECULAR MICROBIOLOGY, 1997, 26 (01) :49-64

[9] Biodiversity and classification of lactococcal phages [J].

Deveau, Helene ;

Labrie, Simon J. ;

Chopin, Marie-Christine ;

Moineau, Sylvain .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (06) :4338-4346

[10] The structure of P4 procapsids produced by coexpression of capsid and external scaffolding proteins [J].

Dokland, T ;

Wang, SF ;

Lindqvist, BH .

VIROLOGY, 2002, 298 (02) :224-231

← 1 2 3 4 5 6 →