Interspecies differences in protein expression do not impact the spatiotemporal regulation of glycoprotein VI mediated activation

Research output: Contribution to journalArticle

Authors

  • Joanne L Dunster
  • Amanda Unsworth
  • Alexander P Bye
  • Marcin A Sowa
  • Ying Di
  • Tanya Sage
  • Chiara Pallini
  • Alexander Hardy
  • Bernhard Nieswandt
  • Angel García
  • Jonathan M Gibbins
  • Alice Y Pollitt

Colleges, School and Institutes

External organisations

  • Institute for Cardiovascular and Metabolic Research (ICMR), School of Biological Sciences, University of Reading, Reading,
  • Department of Experimental Biomedicine; University Hospital, University of Würzburg; Würzburg Germany
  • Institute for Cardiovascular and Metabolic Research, Harborne Building, University of Reading, Reading

Abstract

Background: Accurate protein quantification is a vital prerequisite for generating meaningful predictions when using systems biology approaches, a method that is increasingly being used to unravel the complexities of subcellular interactions and as part of the drug discovery process. Quantitative proteomics, flow cytometry, and western blotting have been extensively used to define human platelet protein copy numbers, yet for mouse platelets, a model widely used for platelet research, evidence is largely limited to a single proteomic dataset in which the total amount of proteins was generally comparatively higher than those found in human platelets. Objectives: To investigate the functional implications of discrepancies between levels of mouse and human proteins in the glycoprotein VI (GPVI) signalling pathway using a systems pharmacology model of GPVI. Methods: The protein copy number of mouse platelet receptors was determined using flow cytometry. The Virtual Platelet, a mathematical model of GPVI signalling, was used to determine the consequences of protein copy number differences observed between human and mouse platelets. Results and conclusion: Despite the small size of mouse platelets compared to human platelets they possessed a greater density of surface receptors alongside a higher concentration of intracellular signalling proteins. Surprisingly the predicted temporal profile of Syk activity was similar in both species with predictions supported experimentally. Super resolution microscopy demonstrates that the spatial distribution of Syk is similar between species, suggesting that the spatial distribution of receptors and signalling molecules in activated platelets, rather than their copy number, is important for signalling pathway regulation.

Details

Original languageEnglish
JournalJournal of Thrombosis and Haemostasis
Early online date4 Nov 2019
Publication statusE-pub ahead of print - 4 Nov 2019

Keywords

  • GPVI, platelet, Syk, mathematical modelling