The precise mechanisms through which post-translational modifications modulate protein function are not clear nor is it understood how they are implicated in human diseases at the molecular level. The objective of this proposal is to develop a method that reveals how variations in sequence and post-translational modifications modulate the structural heterogeneity of proteins. Guided by our strong preliminary data, we will obtain the objective of this proposal by pursuing the following specific aims: 1) To enable structure-elucidation of differentially modified proteins by ion mobility spectrometry / mass spectrometry; and 2) To characterize the structural heterogeneity of differentially modified proteins by tandem-trapped ion mobility spectrometry / mass spectrometry. In the first Aim, we will develop a method that determines structures for differentially modified proteins and their assemblies, in particular for phosphorylated and glycosylated species. In the second Aim, we will develop an approach that reveals how protein structure depends on amino acid sequence and post- translational modifications. The research proposed in this application is innovative because it substantially advances from the status quo through unique computational and experimental methods that were recently developed in our lab, namely the Structure Relaxation Approximation and tandem-trapped ion mobility spectrometry/mass spectrometry methods. This contribution is significant because it is the first step towards a general analytical method that is expected to provide a molecular-level understanding of how changes in amino acid sequence and post-translational modifications are implicated in disease mechanisms. Ultimately, the results of the proposed work can be expected to significantly benefit a number of research areas relevant to the mission of the NIH, including the development of a vaccine against HIV/AIDS.

Public Health Relevance

The proposed research is relevant to public health because a method that is able to link the structural heterogeneity of protein systems to the presence of variations in amino acid sequence and post- translational modifications is ultimately expected to lead to new approaches to preventive and therapeutic purposes, in particular to the development of a vaccine against HIV/AIDS. The proposed research is relevant to the part of NIH's mission that pertains to developing fundamental knowledge that enhances human health.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM135682-01
Application #
9867093
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Smith, Ward
Project Start
2019-09-20
Project End
2023-07-31
Budget Start
2019-09-20
Budget End
2020-07-31
Support Year
1
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Florida State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
790877419
City
Tallahassee
State
FL
Country
United States
Zip Code
32306