TITLE:

GOING LATIN USING IN SILICO, IN VITRO AND IN VIVO METHODS TO FULLY CHARACTERIZE PROTEIN STABILITY/FLEXIBILITY (No. 118)

DATE:

Friday, August 25th, 2006

TIME:

3:30 PM

LOCATION:

GMCS 214

SPEAKER:

John Love, Department of Chemistry, San Diego State University

ABSTRACT:

The correlation between protein structure and function is well established yet the role flexibility plays in protein function is still currently being explored. Here we describe an in vivo screen in which the thermal stability and conformational specificity of a series of test proteins are directly correlated to transcriptional regulation. The system entails the use of a chimeric protein construct that consists of three covalently linked domains which includes a constant N-terminal DNA binding domain, a variable central test protein, and a constant C-terminal transcriptional activation domain. The ten test proteins are mutant variants of the β1 domain of streptococcal protein-G that fairly evenly span a thermal stability range from as low as 38� C to greater than 100� C. When the chimeric construct contains a test variant of high conformational flexibility the reporter gene is upregulated to a greater extent relative to more stable/less flexible variants. In silico (molecular dynamics and computational design) and in vitro (spectroscopic) methods were combined with this in vivo screen to fully characterize the thermal stability and conformational properties of the ten Gβ1 variants.

HOST:

Jose Castillo

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