Monitoring of small conformational changes by high-precision measurements of hydrodynamic radius with 2-focus fluorescence correlation spectroscopy (2fFCS)

Thomas Dertinger; Iris von der Hocht; Ingo Gregor; Konstantin Komolov; Karl-Wilhelm Koch; Jörg Enderlein

doi:10.1117/12.704930

14 February 2007 Monitoring of small conformational changes by high-precision measurements of hydrodynamic radius with 2-focus fluorescence correlation spectroscopy (2fFCS)

Thomas Dertinger, Iris von der Hocht, Ingo Gregor, Konstantin Komolov, Karl-Wilhelm Koch, Jörg Enderlein

Author Affiliations +

Proceedings Volume 6444, Ultrasensitive and Single-Molecule Detection Technologies II; 64440H (2007) https://doi.org/10.1117/12.704930
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

We report on our application of a new fluorescence-correlation spectroscopy technique, 2-focus FCS, for measuring the hydrodynamic radius of molecules with sub- Ångstrøm precision. The method is applied of monitoring conformational changes of proteins upon ion binding. In particular, we present measurements on Ca²⁺-binding of recoverin. Recoverin belongs to the superfamily of EF-hand Ca²⁺-binding proteins and operates as a Ca²⁺-sensor in vertebrate photoreceptor cells, where it regulates the activity of rhodopsin kinase GRK1 in a Ca²⁺-dependent manner. The protein undergoes conformational changes upon Ca²⁺-changes that are reflected as changes in their hydrodynamic radius. By using 2fFCS we were able to resolve hydrodynamic radius changes of ca. one Ångstrøm and used the Ca²⁺ dependence of this radius for recording binding curves in solution. We compare our results with those obtained by other techniques.

Citation Download Citation

Thomas Dertinger, Iris von der Hocht, Ingo Gregor, Konstantin Komolov, Karl-Wilhelm Koch, and Jörg Enderlein "Monitoring of small conformational changes by high-precision measurements of hydrodynamic radius with 2-focus fluorescence correlation spectroscopy (2fFCS)", Proc. SPIE 6444, Ultrasensitive and Single-Molecule Detection Technologies II, 64440H (14 February 2007); https://doi.org/10.1117/12.704930

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
7 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Calcium

Fluorescence correlation spectroscopy

Proteins

Luminescence

Prisms

Pulsed laser operation

Beam splitters

Show All Keywords

Keywords/Phrases

Search In:

Publication Years