Association of Acinetobacter baumannii EF-Tu with Cell Surface, Outer Membrane Vesicles, and Fibronectin

被引:49
作者
Dallo, Shatha F. [1 ]
Zhang, Bailin [2 ]
Denno, James [3 ]
Hong, Soonbae [1 ]
Tsai, Anyu [1 ]
Haskins, Williams [1 ,4 ,5 ,6 ,7 ,8 ]
Ye, Jing Yong [2 ]
Tao Weitao [1 ,9 ]
机构
[1] Univ Texas San Antonio, Dept Biol, San Antonio, TX 78249 USA
[2] Univ Texas San Antonio, Dept Biomed Engn, San Antonio, TX 78249 USA
[3] Univ Texas Austin, Dept Biol, Austin, TX 78712 USA
[4] Univ Texas San Antonio, Pediat Biochem Lab, San Antonio, TX 78249 USA
[5] Univ Texas San Antonio, Dept Chem, San Antonio, TX 78249 USA
[6] Univ Texas San Antonio, RCMI Prote & Prot Biomar Feers Cores, San Antonio, TX 78249 USA
[7] Univ Texas San Antonio, Ctr Res & Training Sci, San Antonio, TX 78249 USA
[8] Univ Texas Hlth Sci Ctr San Antonio, Canc Therapy & Res Ctr, Dept Med, Div Hematol Oncol, San Antonio, TX 78229 USA
[9] SW Baptist Univ, Coll Sci & Math, Dept Biol, Bolivar, MO 65613 USA
关键词
ELONGATION-FACTOR-TU; PHOTONIC CRYSTAL-STRUCTURE; TOTAL-INTERNAL-REFLECTION; NEISSERIA-GONORRHOEAE; ESCHERICHIA-COLI; POLYACRYLAMIDE-GELS; PROTEOMIC ANALYSIS; BINDING PROTEINS; SEROGROUP-B; BACTERIAL;
D O I
10.1100/2012/128705
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A conundrum has long lingered over association of cytosol elongation factor Tu (EF-Tu) with bacterial surface. Here we investigated it with Acinetobacter baumannii, an emerging opportunistic pathogen associated with a wide spectrum of infectious diseases. The gene for A. baumannii EF-Tu was sequenced, and recombinant EF-Tu was purified for antibody development. EF-Tu on the bacterial surface and the outer membrane vesicles (OMVs) was revealed by immune electron microscopy, and its presence in the outer membrane (OM) and the OMV subproteomes was verified by Western blotting with the EF-Tu antibodies and confirmed by proteomic analyses. EF-Tu in the OM and the OMV subproteomes bound to fibronectin as detected by Western blot and confirmed by a label-free real-time optical sensor. The sensor that originates from photonic crystal structure in a total-Internal-reflection (PC-TIR) configuration was functionalized with fibronectin for characterizing EF-Tu binding. Altogether, with a novel combination of immunological, proteomical, and biophysical assays, these results suggest association of A. baumannii EF-Tu with the bacterial cell surface, OMVs, and fibronectin.
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页数:10
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