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

Specificity determinants for lipids bound to beta-barrel proteins.

Amy J Reese1, Leonard J Banaszak

  • 1Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO, USA.

Journal of Lipid Research
|November 5, 2003
PubMed
Summary

Altering a key protein loop in adipocyte lipid-binding protein (ALBP) changed how it binds fatty acids. This structural modification impacts ligand positioning and protein stability, offering insights into lipid-binding protein function.

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

An Undergraduate Elective Course That Introduces Topics of Diversity, Equity, and Inclusion into Discussions of Science.

Journal of microbiology & biology education·2020
Same author

Cell Envelope Integrity and Capsule Characterization of Rhodotorula mucilaginosa Strains from Clinical and Environmental Sources.

mSphere·2019
Same author

X-ray crystallographic analysis of adipocyte fatty acid binding protein (aP2) modified with 4-hydroxy-2-nonenal.

Protein science : a publication of the Protein Society·2010
Same author

The assembly of apoB-containing lipoproteins: a structural biology point of view.

Annals of medicine·2008
Same author

Loss of cell wall alpha(1-3) glucan affects Cryptococcus neoformans from ultrastructure to virulence.

Molecular microbiology·2007
Same author

Vmax regulation through domain and subunit changes. The active form of phosphoglycerate dehydrogenase.

Biochemistry·2005

Area of Science:

  • Structural biology
  • Protein biochemistry
  • Lipid metabolism

Background:

  • Intracellular lipid transport proteins share a conserved 10-stranded beta-barrel structure with an internal binding cavity.
  • Conformational differences in loop regions, specifically the E-F loop, distinguish fatty acid-binding proteins (e.g., adipocyte lipid-binding protein, ALBP) from retinoid-binding proteins (e.g., cellular retinoic acid-binding protein I, CRABP I).

Purpose of the Study:

  • To investigate the functional impact of altering the E-F loop in ALBP to mimic CRABP I.
  • To analyze the structural consequences of this mutation on ligand binding and protein conformation using X-ray crystallography.

Main Methods:

  • Site-directed mutagenesis was employed to introduce a three-residue mutation into wild-type ALBP (WT-ALBP) to create EF-ALBP.

Related Experiment Videos

  • X-ray crystallography was used to determine the crystal structures of ligand-free EF-ALBP and EF-ALBP bound to oleic acid at 1.5 Å and 1.7 Å resolution, respectively.
  • Comparative analysis of the mutant structures with previously determined WT-ALBP structures.

    • Main Results:

    • The three-residue mutation in the E-F loop of ALBP altered the positioning of the bound C18 fatty acid (oleic acid) within the binding cavity, as evidenced by electron density.
    • Crystallographic data allowed for direct comparison of protein conformation and ligand positioning between EF-ALBP and WT-ALBP.
    • Preliminary chemical characterization revealed alterations in the binding properties and overall stability of the mutant EF-ALBP protein.

    Conclusions:

    • Modifications in a single loop region of ALBP can significantly influence fatty acid binding and positioning.
    • Structural insights from EF-ALBP suggest that ligand atoms tend to occupy planar regions within the binding cavities of both fatty acid and retinoid binding proteins.
    • The study provides a structural basis for understanding how subtle conformational changes affect the function and stability of lipid-binding proteins.