Grant: Research Tools and Instruments - Category 1 (<$150,000)
University of Western Ontario
$149,957.00
An Integrated System for Studying Protein Structure and Dynamics by Hydrogen Exchange Mass Spectrometry
EF 2011-12
Résumé
This proposal is for a nanoACQUITY/HDX system that is required for studying protein structure and dynamics by mass spectrometry (MS). The cellular functions of proteins are intimately linked to their structures. However, proteins also have to undergo a multitude of conformational motions to perform their biological tasks. This structure/function/dynamics triad remains poorly understood. Pioneering work by Konermann and others has revealed that hydrogen/deuterium exchange (HDX) MS is a powerful tool for addressing this problem. Protein regions that are highly flexible undergo rapid HDX. Isotope exchange is much slower in rigid segments. The HDX workflow involves digestion and subsequent peptide analysis by liquid chromatography/MS under quench conditions. The deuteration kinetics of individual protein segments can be deduced from the resulting mass profiles. The recent development of a commercial HDX-MS platform has dramatically changed the playing field. Several US labs already purchased this nanoACQUITY/HDX technology, but none of these systems are available in Canada yet. This new platform provides greatly improved reproducibility, digestion efficiency, robustness, and software ease-of-use compared to conventional home-built systems. The proposed equipment will allow a significant expansion of Konermann's research program into the area of transmembrane proteins, many of which represent important drug targets. Specifically, the nanoACQUITY/HDX will allow studies on transmembrane permeation, the mechanism of molecular motors (with Dunn), as well as the structural characterization of bacterial membrane enzymes (with Valvano). Using funds obtained elsewhere, Konermann was able to purchase a state-of-the-art Synapt mass spectrometer that represents one half of the HDX platform. The nanoACQUITY/HDX represents the second half that is still missing (for liquid handling/digestion/chromatography). The current proposal offers superb opportunities for HQP training. Failure to secure funding for this equipment would be a severe setback for the applicants' research programs.
Programme
Research Tools and Instruments - Category 1 (<$150,000)
Bénéficiaire
Emplacement
Ontario, Canada
Type de bénéficiaire
Academia
Type
Grant
Source
NSERC's Awards Data
Tel que publié
Application id
422442-2012
Name nom
Konermann, Lars
Department département
Chemistry
Organization id
36
Institution établissement
University of Western Ontario
Province en
Ontario
Province fr
Ontario
Country en
CANADA
Country fr
CANADA
Fiscal year exercice financier
2011
Competition year année de concours
2012
Award amount
149957
Program id
EQPEQ
Program nane en
Research Tools and Instruments - Category 1 (<$150,000)
Program name fr
Outils et instruments de recherche - catégorie 1 (<150 000$)
Group en
Discovery grants programs
Group fr
Programmes de subventions à la découverte
Committee code
1504
Committee name en
Chemistry
Committee name fr
Chimie
Area of application code
1200
Area of application group en
Advancement of knowledge
Area of application group fr
Avancement des connaissances
Area of application en
Advancement of knowledge
Area of application fr
Avancement des connaissances
Research subject code
3404
Research subject group en
Physical chemistry
Research subject group fr
Chimie physique
Research subject en
Biophysical chemistry
Research subject fr
Chimie biophysique
Partie
1
Nb partie
1
Application title
An Integrated System for Studying Protein Structure and Dynamics by Hydrogen Exchange Mass Spectrometry
Keyword
NSERC
Application summary
This proposal is for a nanoACQUITY/HDX system that is required for studying protein structure and dynamics by mass spectrometry (MS). The cellular functions of proteins are intimately linked to their structures. However, proteins also have to undergo a multitude of conformational motions to perform their biological tasks. This structure/function/dynamics triad remains poorly understood. Pioneering work by Konermann and others has revealed that hydrogen/deuterium exchange (HDX) MS is a powerful tool for addressing this problem. Protein regions that are highly flexible undergo rapid HDX. Isotope exchange is much slower in rigid segments. The HDX workflow involves digestion and subsequent peptide analysis by liquid chromatography/MS under quench conditions. The deuteration kinetics of individual protein segments can be deduced from the resulting mass profiles. The recent development of a commercial HDX-MS platform has dramatically changed the playing field. Several US labs already purchased this nanoACQUITY/HDX technology, but none of these systems are available in Canada yet. This new platform provides greatly improved reproducibility, digestion efficiency, robustness, and software ease-of-use compared to conventional home-built systems. The proposed equipment will allow a significant expansion of Konermann's research program into the area of transmembrane proteins, many of which represent important drug targets. Specifically, the nanoACQUITY/HDX will allow studies on transmembrane permeation, the mechanism of molecular motors (with Dunn), as well as the structural characterization of bacterial membrane enzymes (with Valvano). Using funds obtained elsewhere, Konermann was able to purchase a state-of-the-art Synapt mass spectrometer that represents one half of the HDX platform. The nanoACQUITY/HDX represents the second half that is still missing (for liquid handling/digestion/chromatography). The current proposal offers superb opportunities for HQP training. Failure to secure funding for this equipment would be a severe setback for the applicants' research programs.
