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Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MSMS Quantification of Hepatic Drug Transporters
Joint Authors
Prasad, Bhagwat
Unadkat, Jashvant D.
Source
International Journal of Proteomics
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-02-25
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Biology
Abstract EN
We studied the precision of quantification of organic anion-transporting polypeptide 1B1 (OATP1B1), OATP1B3, OATP2B1, and P-glycoprotein (P-gp) in human livers by surrogate peptide based LC-MS/MS approach using two different internal standards: stable isotope labeled peptide (SIL) versus stable isotope labeled protein (SILAC).
The SIL peptides were procured commercially and the SILAC proteins were generated in-house by labeling arginine and/or lysine residues in cells expressing these transporters.
Liver tissue n=20 was homogenized and the membrane fraction was isolated.
The membranes were trypsin digested and the peptides were analyzed using LC-MS/MS under optimized conditions.
The precision in the quantification of proteins in three independently trypsin digested samples from each liver was calculated as the standard deviation of the log transformed protein concentration.
The precision of the SIL internal standard method was either slightly (P<0.05, paired t-test) better than that of the SILAC method (OATP1B1, OATP1B3, and P-gp) or not different (OATP2B1).
Trypsin digestion, as measured by the response of the labeled peptide derived from the SILAC protein, was consistent across liver samples.
These results indicate that when maximum trypsin digestion is ensured, the SIL internal standard method can be used with confidence for quantification of drug transporters.
American Psychological Association (APA)
Prasad, Bhagwat& Unadkat, Jashvant D.. 2014. Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MSMS Quantification of Hepatic Drug Transporters. International Journal of Proteomics،Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-472628
Modern Language Association (MLA)
Prasad, Bhagwat& Unadkat, Jashvant D.. Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MSMS Quantification of Hepatic Drug Transporters. International Journal of Proteomics No. 2014 (2014), pp.1-11.
https://search.emarefa.net/detail/BIM-472628
American Medical Association (AMA)
Prasad, Bhagwat& Unadkat, Jashvant D.. Comparison of Heavy Labeled (SIL) Peptide versus SILAC Protein Internal Standards for LC-MSMS Quantification of Hepatic Drug Transporters. International Journal of Proteomics. 2014. Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-472628
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-472628