The alterations of mouse plasma proteins during septic development.
A fundamental issue for sepsis therapy is to control the development of inflammation at an early stage. With cecal ligation and puncture (CLP) surgery, the mouse model has clearly shown the septic signs triggered by chronic insult. To monitor the plasma proteomic responses to sepsis, the mouse blood was collected at intervals after sham and CLP surgery followed by the sample treatment to remove high abundance serum albumin. The treated mouse plasma proteins were well resolved by two-dimensional electrophoresis (2-DE). The image analysis revealed that these 2-DE spots observed from the sham and the CLP samples 4 h after surgery were comparable, whereas more than 30 different spots appeared on the 2-DE gels between the sham and CLP mouse plasma 24 h after surgery, indicating that some plasma proteins responded to the inflammatory development. These differential spots were verified by MALDI-TOF/TOF MS, resulting in 13 unique sepsis-responsive proteins. More importantly, most of them exhibited multiple spots as difference on the 2-DE gels. Furthermore, these isospots were incubated with PNGase F to eliminate N-linked oligosaccharides on proteins and then evaluated by Western blot as well as mass spectrometry. The results of PNGase F digestion suggested that most sepsis-associated proteins remained in N-glycosylation status but changed their N-glycans during septic development.
Ren
Yan
Y
Beijing Genomics Institute, Chinese Academy of Science, Beijing, China.
Wang
Jiandong
J
Xia
Ji
J
Jiang
Chaoguang
C
Zhao
Kang
K
Li
Rong
R
Xu
Ningzhi
N
Xu
Yingxin
Y
Liu
Siqi
S
D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000032:analysis* / Q000737:chemistry; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D006031:Glycosylation; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry; D040901:Proteomics; D018805:Sepsis / Q000097:blood*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001798:Blood Proteins; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase