Membrane subproteomic analysis of Mycobacterium bovis bacillus Calmette-Guérin.
Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine has been known for a long time to prevent tuberculosis (TB) worldwide since 1921. Nonetheless, we know little about BCG membrane proteome. In the present study, we utilized alkaline incubation and Triton X-114-based methods to enrich BCG membrane proteins and subsequently digested them using proteolytic enzyme. The recovered peptides were further separated by 2-D LC and identified by ESI-MS/MS. As a result, total 474 proteins were identified, including 78 integral membrane proteins (IMPs). Notably, 18 BCG IMPs were described for the first time in mycobacterium. Further analysis of the 78 IMPs indicated that the theoretical molecular mass distribution of them ranged from 8.06 to 167.86 kDa and pI scores ranged from 4.40 to 11.60. Functional classification revealed that a large proportion of the identified IMPs (67.9%, 53 out of 78) were involved in cell wall and cell processes functional group. In conclusion, here we reported a comprehensive profile of the BCG membrane subproteome. The present investigation may allow the identification of some valuable vaccine and drug target candidates and thus provide basement for future designing of preventive, diagnostic, and therapeutic strategies against TB.
Zheng
Jianhua
J
College of Biological Sciences, China Agricultural University, Beijing, PR China.
Wei
Candong
C
Leng
Wenchuan
W
Dong
Jie
J
Li
Rui
R
Li
Weijun
W
Wang
Jin
J
Zhang
Zongde
Z
Jin
Qi
Q
D000595:Amino Acid Sequence; D001500:BCG Vaccine / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D002853:Chromatography, Liquid; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007526:Isoelectric Point; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009163:Mycobacterium bovis / Q000737:chemistry* / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D011770:Pyruvate Kinase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D053719:Tandem Mass Spectrometry; D014376:Tuberculosis / Q000517:prevention & control
D001500:BCG Vaccine; D001426:Bacterial Proteins; D002352:Carrier Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D011770:Pyruvate Kinase