Differential binding of calmodulin-related proteins to their targets revealed through high-density Arabidopsis protein microarrays.

@article{citeulike:1357061, abstract = {Calmodulins (CaMs) are the most ubiquitous calcium sensors in eukaryotes. A number of CaM-binding proteins have been identified through classical methods, and many proteins have been predicted to bind CaMs based on their structural homology with known targets. However, multicellular organisms typically contain many CaM-like (CML) proteins, and a global identification of their targets and specificity of interaction is lacking. In an effort to develop a platform for large-scale analysis of proteins in plants we have developed a protein microarray and used it to study the global analysis of CaM/CML interactions. An Arabidopsis thaliana expression collection containing 1,133 ORFs was generated and used to produce proteins with an optimized medium-throughput plant-based expression system. Protein microarrays were prepared and screened with several CaMs/CMLs. A large number of previously known and novel CaM/CML targets were identified, including transcription factors, receptor and intracellular protein kinases, F-box proteins, RNA-binding proteins, and proteins of unknown function. Multiple CaM/CML proteins bound many binding partners, but the majority of targets were specific to one or a few CaMs/CMLs indicating that different CaM family members function through different targets. Based on our analyses, the emergent CaM/CML interactome is more extensive than previously predicted. Our results suggest that calcium functions through distinct CaM/CML proteins to regulate a wide range of targets and cellular activities.}, address = {Department of *Molecular, Cellular, and Developmental Biology, Yale University, 219 Prospect Street, New Haven, CT 06520-8103, USA.}, author = {Popescu, S. C. and Popescu, G. V. and Bachan, S. and Zhang, Z. and Seay, M. and Gerstein, M. and Snyder, M. and Dinesh-Kumar, S. P. }, citeulike-article-id = {1357061}, doi = {10.1073/pnas.0611615104}, issn = {0027-8424}, journal = {Proc Natl Acad Sci U S A}, keywords = {calcium}, month = {March}, number = {11}, pages = {4730--4735}, posted-at = {2007-06-02 13:27:10}, priority = {2}, title = {Differential binding of calmodulin-related proteins to their targets revealed through high-density Arabidopsis protein microarrays.}, url = {http://dx.doi.org/10.1073/pnas.0611615104}, volume = {104}, year = {2007} }

TY - JOUR ID - citeulike:1357061 L3 - citeulike-article-id:1357061 TI - Differential binding of calmodulin-related proteins to their targets revealed through high-density Arabidopsis protein microarrays. JF - Proc Natl Acad Sci U S A VL - 104 IS - 11 SP - 4730 EP - 4735 AD - Department of *Molecular, Cellular, and Developmental Biology, Yale University, 219 Prospect Street, New Haven, CT 06520-8103, USA. SN - 0027-8424 N2 - Calmodulins (CaMs) are the most ubiquitous calcium sensors in eukaryotes. A number of CaM-binding proteins have been identified through classical methods, and many proteins have been predicted to bind CaMs based on their structural homology with known targets. However, multicellular organisms typically contain many CaM-like (CML) proteins, and a global identification of their targets and specificity of interaction is lacking. In an effort to develop a platform for large-scale analysis of proteins in plants we have developed a protein microarray and used it to study the global analysis of CaM/CML interactions. An Arabidopsis thaliana expression collection containing 1,133 ORFs was generated and used to produce proteins with an optimized medium-throughput plant-based expression system. Protein microarrays were prepared and screened with several CaMs/CMLs. A large number of previously known and novel CaM/CML targets were identified, including transcription factors, receptor and intracellular protein kinases, F-box proteins, RNA-binding proteins, and proteins of unknown function. Multiple CaM/CML proteins bound many binding partners, but the majority of targets were specific to one or a few CaMs/CMLs indicating that different CaM family members function through different targets. Based on our analyses, the emergent CaM/CML interactome is more extensive than previously predicted. Our results suggest that calcium functions through distinct CaM/CML proteins to regulate a wide range of targets and cellular activities. KW - calcium AU - Popescu, SC AU - Popescu, GV AU - Bachan, S AU - Zhang, Z AU - Seay, M AU - Gerstein, M AU - Snyder, M AU - Dinesh-Kumar, SP PY - 2007/03/13/ UR - http://dx.doi.org/10.1073/pnas.0611615104 ER -