EXROP Projects: John Kuriyan

John Kuriyan
Summer Lab Size: 20
Local Summer Program: Berkeley SROP/AMGEN Program
Program Dates: June 1-August 7, 2015 (Dates for 2016 should be similar)

Structural and Biochemical Studies of Ca2+/Calmodulin-Dependent Protein Kinase II (CaMKII)

CaMKII is a highly conserved, multimeric kinase that represents 1% of your dry brain weight. It plays a key role in mammalian learning and memory. Upon activation, CaMKII holoenzymes can exchange subunits, resulting in a spread of kinase activity. The ultimate goal of this project is to understand CamKII’s subunit exchange at a molecular level. Employed techniques will include: x-ray crystallography, native protein mass spectrometry, fluorescence resonance energy transfer spectroscopy, analytical gel filtration chromatography, molecular cloning, and heterologous protein expression and purification.

High-Throughput Approach to Understanding B and T Cell Kinase Specificity

The tyrosine kinase ZAP-70 plays a crucial role in T cell receptor (TCR) signaling. ZAP-70 is unique among tyrosine kinases, including the paralogous B cell kinase Syk, in that it has very narrow substrate specificity. Although this specificity is required for stringent control of TCR signaling, it is not clear how ZAP-70 recognizes its substrates. To understand the structural basis of this specificity, we will use chemically synthesized peptide-ATP conjugates to obtain co-crystals and solve high-resolution complex structures. In addition, we have developed high-throughput methods to simultaneously determine the activity of ZAP-70 towards thousands of substrate mutants.

Investigation of the RING-Type E3 Ligase Cbl From Multiple Organisms

This project involves understanding the relationship between tyrosine kinases and Cbl  an E3 ligase that ubiquitylates these kinase substrates, targeting them for degradation. Specifically, we are interested in the protein-protein interactions of tyrosine kinases and the tyrosine kinase-binding domain of Cbl homologues from distantly related organisms (e.g., D. discoideum, S. rosetta, and C. elegans). In this project, we will use in vitro biochemical assays, enzyme kinetics, and x-ray crystallography.

Scientist Profile

Investigator
University of California, Berkeley
Biophysics, Structural Biology