Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Mapping global angular transitions of proteins in assemblies using multiple extrinsic reporter groups.

T P Burghardt1, K Ajtai

  • 1Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, Minnesota 55905.

Biochemistry
|January 14, 1992
PubMed
Summary

This study introduces a model-independent method to determine the orientation and order parameters of molecular probes in biological assemblies like muscle fibers. The technique reveals myosin SH1 rotation during nucleotide binding, primarily a torsional motion.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

In vitro actin motility velocity varies linearly with the number of myosin impellers.

Archives of biochemistry and biophysics·2017
Same author

Myosin head rotation in muscle fibers measured using polarized fluorescence photobleaching recovery.

Journal of fluorescence·2013
Same author

Rotations of a few cross-bridges in muscle by confocal total internal reflection microscopy.

Biochimica et biophysica acta·2006
Same author

Changes in orientation of actin during contraction of muscle.

Biophysical journal·2004
Same author

Structural characterization of beta-cardiac myosin subfragment 1 in solution.

Biochemistry·2001
Same author

Effect of ionic strength on the conformation of myosin subfragment 1-nucleotide complexes.

Biophysical journal·2001

Area of Science:

  • Biophysics
  • Structural Biology
  • Biochemistry

Background:

  • Understanding molecular dynamics in biological systems is crucial for deciphering cellular functions.
  • Probes like fluorescent and spin labels are used to study molecular motion, but their interpretation can be complex.

Purpose of the Study:

  • To develop a model-independent analytical method for determining probe order parameters and relative orientations.
  • To investigate the rotational dynamics of myosin sulfhydryl 1 (SH1) in muscle fibers upon nucleotide binding.

Main Methods:

  • Utilized fluorescence polarization and electron paramagnetic resonance spectroscopy to obtain probe order parameters.
  • Developed a system of equations based on Euler rotations to link order parameters from different probes.
  • Applied a model-independent approach to analyze experimental data without prior assumptions.

Related Experiment Videos

Main Results:

  • Successfully determined Euler angles representing the relative orientation of probe molecular frames.
  • Obtained a comprehensive set of probe order parameters exceeding direct experimental resolution.
  • Demonstrated that the myosin SH1 transition from rigor to MgADP binding is mainly a rotation about the cross-bridge's axis of symmetry.

Conclusions:

  • The developed analytical method provides high-resolution insights into molecular orientations and dynamics.
  • The findings suggest a torsional mechanism for the myosin cross-bridge conformational change during nucleotide interaction.