Optimization of the SDS-PAGE gel slicing approach for identification of human liver microsomal proteins via MALDI-TOF mass spectrometry
- *Corresponding Author:
- Natalia Petushkova
Institute of Biomedical Chemistry
Russian Academy of Medical Sciences, 119121
Pogodinskaya St., 10, Moscow, Russia
Tel: +7 (499) 255- 39-60
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E-mail: [email protected], [email protected]
Received Date: December 21, 2011; Accepted Date: January 21, 2012; Published Date: January 31, 2012
Citation: Petushkova NA, Larina OV, Chernobrovkin AL, Trifonova OP, Kisrieva YS, et al. (2012) Optimization of the SDS-PAGE gel slicing approach for identification of human liver microsomal proteins via MALDI-TOF mass spectrometry. J Proteomics Bioinform 5: 040-049. doi: 10.4172/jpb.1000211
Copyright: © 2012 Petushkova NA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Proteomics often exploits sequential cutting of the SDS-PAGE gel lane into slices with subsequent identification of proteins by peptide mass fingerprinting (PMF). In this paper, the influence of slice thickness on protein identification was investigated. Following the separation of the human liver microsomal fraction, the 37 to 75 kDa range of the gel lane was cut into 20 or 40 slices of 0.5 mm and 0.25 mm thickness, respectively. Identification of proteins after trypsinolysis was performed by MALDI-TOF mass spectrometry. A twofold reduction of slice thickness did not influence the number of peaks in PMF-spectra. It was established that at a tolerance of 0.15 Da the number of identified proteins in the 40 slices series exceeded by more than twofold the number of identifications in 20 slices series. It was shown that the decrease of slice thickness leads to a considerable enhancement of peptide peak’s intensity and, also, to the changes of PMF-spectra which resulted gaining additional peptides for enrichment of sequence coverage.