Proteomic and computational analysis of secreted proteins with type 1 signal peptides from the antarctic archaeon Methanococcoides burtonii

被引:29
作者
Saunders, Neil F. W.
Ng, Charmaine
Raftery, Mark
Guilhaus, Michael
Goodchild, Amber
Cavicchioli, Ricardo [1 ]
机构
[1] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia
[2] Univ New S Wales, Bioanalyt Mass Spect Facil, Sydney, NSW 2052, Australia
[3] Johnson & Johnson Res, Eveleigh, NSW 1430, Australia
关键词
LC-MS/MS; secretion; supernatant fraction; type 1 signal peptides; archaea; cold adaptation; psychrophile; protease; S-layer protein; SignaIP;
D O I
10.1021/pr060220x
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
LC-MS/MS was used to identify secreted proteins in the Antarctic archaeon Methanococcoides burtonii. Seven proteins possessing a classical class 1 signal peptide were identified in the supernatant from cultures grown at 4 and 23 C. The proteins included a putative S-layer cell surface protein, cell surface protein involved with cell adhesion, and trypsin-like serine protease. Protease activity was detected in the secreted fraction, and the signal peptide cleavage site of the protease was confirmed using Edman sequencing. The expression profile of putative cell surface proteins suggests a requirement for cell interactions during growth at low temperature. Sequences of the secreted proteins were used to compile a dataset containing a further 32 predicted secreted proteins from the Methanosarcinaceae. Many of these proteins were also S-layer cell surface proteins with a variety of predicted roles, particularly in cell-cell interaction. Computational analysis of signal peptides revealed a preference for lysine in the n-region, leucine in the h-region, and a eucaryal-type cleavage site, highlighting the mosaic nature of signal peptides in Archaea. This is the first study to experimentally characterize secreted proteins from a cold-adapted archaeon and provides new insight and a functional dataset for studying secretion in Archaea.
引用
收藏
页码:2457 / 2464
页数:8
相关论文
共 46 条
[1]   Cleavage of preflagellins by an aspartic acid signal peptidase is essential for flagellation in the archaeon Methanococcus voltae [J].
Bardy, SL ;
Jarrell, KF .
MOLECULAR MICROBIOLOGY, 2003, 50 (04) :1339-1347
[2]   Archaeal signal peptides - A comparative survey at the genome level [J].
Bardy, SL ;
Eichler, J ;
Jarrell, KF .
PROTEIN SCIENCE, 2003, 12 (09) :1833-1843
[3]   Improved prediction of signal peptides: SignalP 3.0 [J].
Bendtsen, JD ;
Nielsen, H ;
von Heijne, G ;
Brunak, S .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 340 (04) :783-795
[4]   IMPROVED SILVER STAINING OF PLANT-PROTEINS, RNA AND DNA IN POLYACRYLAMIDE GELS [J].
BLUM, H ;
BEIER, H ;
GROSS, HJ .
ELECTROPHORESIS, 1987, 8 (02) :93-99
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   Cold-adapted archaea [J].
Cavicchioli, R .
NATURE REVIEWS MICROBIOLOGY, 2006, 4 (05) :331-343
[7]   Low-temperature extremophiles and their applications [J].
Cavicchioli, R ;
Siddiqui, KS ;
Andrews, D ;
Sowers, KR .
CURRENT OPINION IN BIOTECHNOLOGY, 2002, 13 (03) :253-261
[8]  
CAVICCHIOLI R, 2006, IN PRESS MICROBIAL P
[9]   IMMUNOCHEMISTRY AND LOCALIZATION OF THE ENZYME DISAGGREGATASE IN METHANOSARCINA-MAZEI [J].
DEMACARIO, EC ;
MACARIO, AJL ;
MOK, T ;
BEVERIDGE, TJ .
JOURNAL OF BACTERIOLOGY, 1993, 175 (10) :3115-3120
[10]   IMPROVED PROCEDURE FOR PROTEIN STAINING IN POLYACRYLAMIDE GELS WITH A NEW TYPE OF COOMASSIE BRILLIANT BLUE [J].
DIEZEL, W ;
HOFMANN, E ;
KOPPERSC.G .
ANALYTICAL BIOCHEMISTRY, 1972, 48 (02) :617-+