X-Ray Footprinting For Structural Biology

Organizers: Corie Ralston and Sayan Gupta

 

ALS Users’ Meeting
Date: Oct 5, 2016
Venue: Lawrence Berkeley National Laboratory, 6-1105

Workshop Description

X-ray Footprinting (radiolytic hydroxyl radical labeling and mass spectrometry) is a technique that provides structural and dynamical information on proteins in the solution state and is highly complementary to many other structural and biochemical techniques. Because it yields residue-specific information by in-situ generation of hydroxyl radicals, it is used to determine conformation changes, ligand binding and protein-protein interactions under various conditions at a high degree of both temporal and spatial resolution and therefore is useful to both fundamental research and characterization of biologics/protein therapeutics. The technique was pioneered at the NSLS a decade ago and is now available at the ALS. This workshop will provide an overview of the X-Ray footprinting technique, how it is performed, the best way to prepare samples, and what information the technique provides. The morning session will highlight recent case studies, and the afternoon session is a tutorial on data-processing. The workshop will be useful for biological researchers in both academia and the pharmaceutical industry.

 

Agenda 

TIME SESSION
9:30-11:30am

Current Applications in X-Ray Footprinting

Overview of X-Ray Footprinting: The Information it Provides

Corie Ralston
Molecular Biophysics and Integrated Bioimaging, LBNL

 

Using Footprinting to Characterize Gating Events in Zinc transporter Proteins

Dax Fu
Johns Hopkins University

 

Characterizing Conformational Changes in Chloride Transporters

Tanmay Chavan
Stanford University

 

The Orange Carotenoid Protein: Mechanism of  a Photoswitch

Corie Ralston
Molecular Biophysics and Integrated Bioimaging, LBNL

 

Designing and Characterizing Organic-IronOxide Interfaces and Applications for Biohybrid Engineering

Behzad Rad
Materials Science Division, LBNL
11:30am-1:00pm LUNCH
1:00-2:00pm

Latest Technique Development

Application of MS-based Footprinting in Drug Discovery and Development

Janna Kiselar
Case Western Reserve University

 

In situ X-ray Protein Footprinting of Intact, Functional Mitochondria

Awuri Asuru
Case Western Reserve University

 

The New X-Ray Footprinting Beamline at the NSLS-II

Slides: 1-10, 11-23

Mark Chance
Case Western Reserve University, School of Medicine Vice Dean for Research

 

2:00-4:00

Hands-On Tutorial

Footprinting Strategies: LC-MS and Data Analysis

Slides: First half, Second half

Sayan Gupta and Janna Kiselar

 4:00-4:30 COFFEE BREAK AND DISCUSSION
 4:30-5:00

Tour of the ALS Footprinting Beamlines