A Strategy of Plasma Protein Quantitation by Selective Reaction Monitoring of an Intact Protein

2003 ◽  
Vol 75 (24) ◽  
pp. 7008-7014 ◽  
Author(s):  
Qin C. Ji ◽  
Ramona Rodila ◽  
Eric M. Gage ◽  
Tawakol A. El-Shourbagy
Keyword(s):  
Plasma Protein ◽  
Intact Protein ◽  
Reaction Monitoring ◽  
Selective Reaction Monitoring ◽  
Selective Reaction ◽  
Protein Quantitation

scholarly journals The application of selective reaction monitoring confirms dysregulation of glycolysis in a preclinical model of schizophrenia

2012 ◽  
Vol 5 (1) ◽  
Author(s):  
Daniel Martins-de-Souza ◽  
Murtada Alsaif ◽  
Agnes Ernst ◽  
Laura W Harris ◽  
Nancy Aerts ◽  
...  
Keyword(s):  
Preclinical Model ◽  
Reaction Monitoring ◽  
Selective Reaction Monitoring ◽  
Selective Reaction

scholarly journals Genomic tagging reveals a random association of endogenous PtdIns5P 4-kinases IIα and IIβ and a partial nuclear localization of the IIα isoform

2010 ◽  
Vol 430 (2) ◽  
pp. 215-221 ◽  
Author(s):  
Minchuan Wang ◽  
Nicholas J. Bond ◽  
Andrew J. Letcher ◽  
Jonathan P. Richardson ◽  
Kathryn S. Lilley ◽  
...  
Keyword(s):  
Cell Signalling ◽  
Internal Standard ◽  
Absolute Amount ◽  
Reaction Monitoring ◽  
Type Ii ◽  
Real Time Quantitative Pcr ◽  
Selective Reaction Monitoring ◽  
Selective Reaction ◽  
Dt40 Cells

PtdIns5P 4-kinases IIα and IIβ are cytosolic and nuclear respectively when transfected into cells, including DT40 cells [Richardson, Wang, Clarke, Patel and Irvine (2007) Cell. Signalling 19, 1309–1314]. In the present study we have genomically tagged both type II PtdIns5P 4-kinase isoforms in DT40 cells. Immunoprecipitation of either isoform from tagged cells, followed by MS, revealed that they are associated directly with each other, probably by heterodimerization. We quantified the cellular levels of the type II PtdIns5P 4-kinase mRNAs by real-time quantitative PCR and the absolute amount of each isoform in immunoprecipitates by MS using selective reaction monitoring with 14N,13C-labelled internal standard peptides. The results suggest that the dimerization is complete and random, governed solely by the relative concentrations of the two isoforms. Whereas PtdIns5P 4-kinase IIβ is >95% nuclear, as expected, the distribution of PtdIns4P 4-kinase IIα is 60% cytoplasmic (all bound to membranes) and 40% nuclear. In vitro, PtdIns5P 4-kinase IIα was 2000-fold more active as a PtdIns5P 4-kinase than the IIβ isoform. Overall the results suggest a function of PtdIns5P 4-kinase IIβ may be to target the more active IIα isoform into the nucleus.

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