Proteomic analysis on the temperature-dependent complexes in Thermoanaerobacter tengcongensis.
It is generally accepted that protein complexes play an active role in avoiding the protein degradation of the thermophiles. Thermoanaerobacter tengcongensis was cultured at three different temperatures (55, 75 and 80 degrees C) and the extracts of protein complexes were prepared. Through blue native PAGE, the changes of the relative band volumes in response to different temperatures were semi-quantitatively compared and six temperature-dependent bands were obtained. These bands were excised, digested with trypsin and then analyzed with MS for the identification of protein components. With the combination of the proteins identified by LC MS/MS and MALDI TOF/TOF MS, a total of 92 unique proteins were ascertained in these complexes. Besides, some protein components were examined with Western blot, which gave us insights into the survival mechanism of thermophiles. These included (i) the composition of complex at 80 degrees C was significantly different from that at the other two temperatures; (ii) HSPs presented in all temperature-dependent complexes; (iii) several proteins associated with the functional pathways existed in the same complexes, indicating that the complex structure provided facility for the functional efficiency.
Meng
Bo
B
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, PR China.
Qian
Zhong
Z
Wei
Fan
F
Wang
Weiwei
W
Zhou
Chuanqi
C
Wang
Zhuowei
Z
Wang
Quanhui
Q
Tong
Wei
W
Wang
Qian
Q
Ma
Yanhe
Y
Xu
Ningzhi
N
Liu
Siqi
S
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D013312:Stress, Physiological; D053719:Tandem Mass Spectrometry; D013696:Temperature; D046968:Thermoanaerobacter / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D001426:Bacterial Proteins; D046912:Multiprotein Complexes