Tissue-specific Identification of CaM Kinase II Binding Partners and Substrates

Virginia Commonwealth University, Richmond VA

Investigators

Abstract

The formation of organisms from a fertilized egg depends on the coordinated expression of genes, whose protein products interact with and influence each other. One of these genes encodes a regulatory protein known as CaM kinase II. This protein is 94% identical between humans and simple vertebrate organisms, such as the zebrafish. In fact, defects in the development of the heart caused by mutations in the zebrafish CaM kinase II gene, can be reversed by introducing the human CaM kinase II protein. This protein normally influences proper heart development by binding to and/or modifying other proteins. Previous studies from this laboratory have used a biochemical technique (peptide mass spectrometry) to identify such protein partners in cells in culture. Proteomic studies like these are ultimately required to define downstream targets and thus functions of regulatory proteins like CaM kinase II. This interdisciplinary project will link molecular tools with proteomics in a whole animal model to identify downstream proteins in specific tissues at any time during development. In particular, such proteins will be identified at specific developmental stages in the embryonic heart. It is anticipated that this project will shed light on the unsolved puzzle of cardiac chamber formation and looping and will begin to identify new binding partners and substrates of the CaM kinase II family. This project will influence our understanding of the development of the cardiovascular system and will develop a template approach for other investigators interested in evaluating tissue-specific changes in the nature of any protein complex during development. Graduate and undergraduate students will be trained.