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

Structural and functional linkages between subunit interfaces in mammalian pyruvate kinase.

J O Wooll1, R H Friesen, M A White

  • 1Department of Human Biological Chemistry and Genetics and the Sealy Center for Structural Biology, The University of Texas Medical Branch, Galveston, TX 77555-0647, USA.

Journal of Molecular Biology
|September 21, 2001
PubMed
Summary

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

Identification of the structural and functional domains of MutY, an Escherichia coli DNA mismatch repair enzyme.

The Journal of biological chemistry·1996
Same author

Human mitogen-activated protein kinase CSBP1, but not CSBP2, complements a hog1 deletion in yeast.

The Journal of biological chemistry·1995
Same author

A CD10-positive subset of malignant cells is identified in multiple myeloma using PCR with patient-specific immunoglobulin gene primers.

Leukemia·1995
Same author

Osteogenic protein-1 enhances phenotypic expression in ROS 17/2.8 cells.

The American journal of physiology·1995
Same author

Identification of a Drosophila muscle development gene with structural homology to mammalian early growth response transcription factors.

Proceedings of the National Academy of Sciences of the United States of America·1995
Same author

Application of ABO genotyping using polymerase chain reaction method to assess engraftment in bone marrow transplantation--a case report.

Gaoxiong yi xue ke xue za zhi = The Kaohsiung journal of medical sciences·1995

Mutating rabbit muscle pyruvate kinase (M1-PK) at the Y interface alters its structure, impacting the Z interface. This reveals how subunit interactions regulate enzyme function and allosteric regulation.

Area of Science:

  • Biochemistry
  • Enzymology
  • Structural Biology

Background:

  • Mammalian pyruvate kinase (PK) exists as tissue-specific isozymes with varying allosteric regulation.
  • Muscle PK (M1-PK) exhibits hyperbolic kinetics, while kidney PK (M2-PK) shows sigmoidal kinetics.
  • Sequence differences between M1-PK and M2-PK are concentrated at inter-subunit interfaces.

Purpose of the Study:

  • To investigate the role of inter-subunit interactions in signal transmission within M1-PK.
  • To understand the functional and structural connectivity between different interfaces in PK.
  • To elucidate how mutations affect allosteric regulation and conformational states.

Main Methods:

  • Crystallization and X-ray diffraction of the S402P mutant of M1-PK.

Related Experiment Videos

  • Structural analysis comparing the mutant to wild-type M1-PK.
  • Comparison with previously resolved PK structures.

    • Main Results:

    • The S402P mutation in M1-PK caused backbone conformational changes at the Y interface.
    • A salt bridge loss at the Z interface (Asp177:Arg341) was observed.
    • Increased rotational flexibility of domain B, containing the active site, correlated with salt bridge loss.

    Conclusions:

    • Structural linkages exist between the Y and Z interfaces in rabbit M1-PK.
    • These linkages mediate the interconversion of enzyme conformational states.
    • Inter-subunit interactions are critical for regulating PK allosteric behavior.