Sample preparation for two-dimensional blue native/SDS polyacrylamide gel electrophoresis in the identification of Streptomyces coelicolor cytoplasmic protein complexes.
Ammonium sulfate precipitation was tested as a sample preparation step for BN-PAGE analyses of S. coelicolor cytoplasmic protein complexes. A procedure of sample preparation compatible with two-dimensional BN/SDS-PAGE was established and used to visualize protein complexes. To validate the sample preparation procedure, representative protein complexes were identified. Several previously characterized protein complexes were rediscovered and their reported oligomeric states reconfirmed. In addition, we identified new but plausible interactions that have never been reported before. Our work provides useful reference for the wide application of BN-PAGE in protein interaction study.
Wang
Zhi-Jun
ZJ
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100080, PR China. zhijunw@gmail.com
Xu
Xin-Ping
XP
Fan
Ke-Qiang
KQ
Jia
Cui-Juan
CJ
Yang
Ke-Qian
KQ
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D003593:Cytoplasm / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification*; D013058:Mass Spectrometry; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013171:Spores, Bacterial / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000502:physiology*; D014357:Trypsin
D001426:Bacterial Proteins; D004798:Enzymes; D014357:Trypsin