Citation: KONG Xiang-Yi, DU Jian-Shi, XU Jin-Ling, LI Shui-Ming, WANG Yong, ZHAO Qing. Comparison of Two Different Separation Methods for Analysis of Salivary Peptidome[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(11): 1816-1822. doi: 10.19756/j.issn.0253-3820.191363 shu

Comparison of Two Different Separation Methods for Analysis of Salivary Peptidome

1.
Lymphatic and Vascular Surgery Department, Xinmin Branch of China Japan Union Hospital, Jilin University, Changchun 130033, China;
2.
College of Life Science and Oceanography, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China;
3.
Neurology Department, Nanhu Branch of China Japan Union Hospital, Jilin University, Changchun 130033, China

Received Date: 27 June 2019
Revised Date: 13 August 2019

Zip-Tip C₁₈ solid phase extraction and oxide graphene-lanthanum phosphate nano composite (LaGM) were used to separate saliva peptides respectively. The peptides were identified by high-resolution tandem time-of-flight mass spectrometry (TOF-MS). To reduce accidental errors, the MS analysis repeated once, and the results of two replicates were combined and then were compared at the peptide and protein levels respectively. A total of 545 sequence-specific peptides belonging to 38 proteins were detected by Zip-Tip C₁₈ method, and 359 different peptides coming from 44 proteins were detected by LaGM method, and the two were coincident with 116 peptides (19 proteins). Furthermore, it was found that the peptide distribution characteristics and the dominant peptide composition obtained by the two methods were significantly different. More peptides of Submaxillary gland androgen-regulated protein 3B, Statherin and Salivary acidic proline-rich phosphoprotein 1/2 were enriched by Zip-Tip C₁₈; more peptides of Histatin-1, Hisatin-3 and Protein S100-A8 were detected by LaGM method, and the conclusions were consistent after post-translational modification were included. This study showed that these two methods had a preference for the enrichment of peptides, e.g. the separation process may lead to the loss of some peptides, the possible missing peptides may be the sequence-related peptides, modification-related peptides, protein-related peptides or peptides coming from some other proteins, fortunately, the potential missing peptides were more likely to be related with the peptides that had been detected. This study suggested that it may be not appropriate to use the results of a single separation method to represent the entire peptidome, and although the LaGM method detected fewer peptides, it was more conducive to enrichment of low-abundance peptides, by which more information about degrading proteins could be obtained.
- Peptididome,
- Zip-Tip C₁₈,
- Oxide graphene-lanthanum phosphate nano composite magnetic beads,
- Saliva,
- High resolution tandem mass spectrometry
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1.
沈阳化工大学材料科学与工程学院沈阳 110142