Intracellular calmodulin availability accessed with two-photon cross-correlation

被引:104
作者
Kim, SA
Heinze, KG
Waxham, MN
Schwille, P
机构
[1] Max Planck Inst Biophys Chem, Expt Biophys Grp, D-3777 Gottingen, Germany
[2] Univ Texas, Hlth Sci Ctr, Dept Neurobiol & Anat, Houston, TX 77030 USA
关键词
D O I
10.1073/pnas.2436461100
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The availability and interactions of signaling proteins are tightly regulated in time and space to produce specific and localized effects. For calmodulin (CaM), a key transducer of intracellular Ca2+ signaling, binding to its variety of targets initiates signaling cascades and regulates its subcellular localization, thereby making it unavailable for subsequent binding interactions. Among CaM's numerous targets, Ca2+ /Ca M-dependent protein kinase 11 is one of the most striking due to its unique ability to increase its affinity for CaM by autophosphorylation and to translocate when bound to Ca2+/CaM. Two-photon fluorescence correlation spectroscopy and cross-correlation spectroscopy were utilized to compare mobility and molecular interactions between CaM and Ca2+/CaM-dependent protein kinase 11 in solution and in living cells. These techniques revealed that CaM availability in cells could be altered by a change in intracellular conditions. Two-photon fluorescence cross-correlation spectroscopy exemplifies a generally applicable approach for studying protein-protein interactions in living cells that allows access to the behavior of signaling molecules within their native environment to probe for heterogeneities in signaling pathways in different cellular compartments.
引用
收藏
页码:105 / 110
页数:6
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