Profiling the secretome of the marine bacterium Pseudoalteromonas tunicata using amine-specific isobaric tagging (iTRAQ)

被引:40
作者
Evans, Flavia F.
Raftery, Mark J.
Egan, Suhelen
Kjelleberg, Staffan [1 ]
机构
[1] Univ New S Wales, Ctr Marine Biofouling & Bioinnovat, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia
[2] Univ New S Wales, Bioanalyt Mass Spect Facil, Sydney, NSW 2052, Australia
关键词
secretome; iTRAQ; LC-MS/MS; GSP; type-II secretion; Pseudoalteromonas sp;
D O I
10.1021/pr060416x
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The eukaryote-associated marine bacterium Pseudoalteromonas tunicata produces a range of target-specific compounds that inhibit different types of marine organisms including invertebrate larvae and algal spores, as well as a broad spectrum of fungi, protozoa, and bacteria. The ability to produce such bioactive compounds is correlated to the expression of a yellow and a purple pigment in P. tunicata. To investigate the regulation and biosynthesis of the pigments and bioactive compounds, the expressed secretome of the pigmented wild-type P. tunicata and a nonpigmented mutant (wmpD(-)) defective in the type-II secretion pathway were compared. Secreted proteins were digested with trypsin, labeled using amine-specific isobaric tagging reagents (iTRAQ), and identified using two-dimensional SCX and nano C18 RP liquid-chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS). The iTRAQ labeling experiments enabled accurate measurement of the proteins identified in this work. A sequence-base prediction of P. tunicata secretome was also obtained and compared to the expressed proteome to determine the role of the type-II secretion pathway in this bacterium. Our results suggest that this secretion pathway has a role in iron transport and acquisition in P. tunicata.
引用
收藏
页码:967 / 975
页数:9
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