# Protein Chemistry & Biophysics Core

> **NIH NIH P01** · UNIVERSITY OF ARIZONA · 2020 · $213,365

## Abstract

ABSTRACT:
 The generation of purified proteins, assessment of protein post-translational modifications (PTMs),
protein-protein interactions and evaluation of cellular mechanical properties are integral to the successful
completion of the Specific Aims proposed in all three PPG projects. The Protein Chemistry & Biophysics Core
will provide services that will support these analyses in all of the PPG Projects. The Specific Aims of Core D
are: 1) Generation of purified recombinant proteins; 2) Analysis of protein PTMs in mechanical stress-
challenged cells and tissues; 3) Analysis of protein-protein interactions; 4) Analysis of changes in
micromechanical properties of endothelial cells (EC) induced by barrier modifying agents. All three
Projects will utilize recombinant proteins for PTM and protein interaction analyses. Proteins generated will be
available for each project and for “in core” use. Additionally, in this aim, proteins from biological samples will be
purified to assay PTMs and protein-protein interactions in vivo. To analyze PTMs, we will utilize a mass
spectrometry approach coupled to the detection and quantification of specific sites of modification by tandem
mass spectrometry. To enhance the analysis of protein-protein interactions (interactome) required in all three
Projects, Core D will utilize surface plasmon resonance (SPR) techniques. In vivo measurements of protein-
protein interactions will also be validated using immunoprecipitation–mass spectrometry identification
approaches. Core D will also utilize atomic force microscopy (AFM) to evaluate structural and elasticity effects
in the endothelium to enable investigators to define the relationship between cell mechanics and cytoskeletal
reorganization in response to mechanical stretch and the effects of barrier protective agents. Core D leaders
have considerable experience with all of the proposed techniques.

## Key facts

- **NIH application ID:** 9959504
- **Project number:** 5P01HL134610-03
- **Recipient organization:** UNIVERSITY OF ARIZONA
- **Principal Investigator:** Stephen M Black
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $213,365
- **Award type:** 5
- **Project period:** — → —

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9959504

## Citation

> US National Institutes of Health, RePORTER application 9959504, Protein Chemistry & Biophysics Core (5P01HL134610-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9959504. Licensed CC0.

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