Conformational changes of calmodulin upon Ca2+ binding studied with a microfluidic mixer.

Abstract:

:A microfluidic mixer is applied to study the kinetics of calmodulin conformational changes upon Ca2+ binding. The device facilitates rapid, uniform mixing by decoupling hydrodynamic focusing from diffusive mixing and accesses time scales of tens of microseconds. The mixer is used in conjunction with multiphoton microscopy to examine the fast Ca2+-induced transitions of acrylodan-labeled calmodulin. We find that the kinetic rates of the conformational changes in two homologous globular domains differ by more than an order of magnitude. The characteristic time constants are approximately 490 micros for the transitions in the C-terminal domain and approximately 20 ms for those in the N-terminal domain of the protein. We discuss possible mechanisms for the two distinct events and the biological role of the stable intermediate, half-saturated calmodulin.

authors

Park HY,Kim SA,Korlach J,Rhoades E,Kwok LW,Zipfel WR,Waxham MN,Webb WW,Pollack L

doi

10.1073/pnas.0710810105

subject

Has Abstract

pub_date

2008-01-15 00:00:00

pages

542-7

issue

2

eissn

0027-8424

issn

1091-6490

pii

0710810105

journal_volume

105

pub_type

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