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

Structure-function studies on human macrophage colony-stimulating factor (M-CSF)

K Koths1

  • 1Chiron Corporation, Emeryville, California 94608-2916, USA.

Molecular Reproduction and Development
|January 1, 1997
PubMed
Summary
This summary is machine-generated.

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

Gelsolin in complex with phosphatidylinositol 4,5-bisphosphate inhibits caspase-3 and -9 to retard apoptotic progression.

The Journal of biological chemistry·2000
Same author

Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis.

Science (New York, N.Y.)·1997
Same author

Interactions between galectin-3 and Mac-2-binding protein mediate cell-cell adhesion.

Cancer research·1996
Same author

The crystal structure of PR3, a neutrophil serine proteinase antigen of Wegener's granulomatosis antibodies.

Journal of molecular biology·1996
Same author

Recombinant proteins for medical use: the attractions and challenges.

Current opinion in biotechnology·1995
Same author

Structure-function studies on recombinant human macrophage colony-stimulating factor (M-CSF).

The Journal of biological chemistry·1994

Recombinant macrophage colony-stimulating factor (rM-CSF) variants, including truncated forms, are functionally equivalent in vitro. Modifying rM-CSF’s structure, like adding polyethylene glycol, significantly enhances its circulating half-life in vivo.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Macrophase colony-stimulating factor (M-CSF) exists in various structural forms impacting in vivo function.
  • Recombinant M-CSF (rM-CSF) from E. coli can be refolded into functional nonglycosylated forms.
  • A conserved N-terminal domain (149 amino acids) is crucial for M-CSF bioactivity.

Purpose of the Study:

  • To investigate the functional equivalence of different M-CSF structural variants.
  • To explore methods for enhancing M-CSF's in vivo half-life.
  • To elucidate the structural basis of M-CSF receptor binding and signaling.

Main Methods:

  • In vitro refolding of truncated and heterodimeric rM-CSF variants.
  • In vivo pharmacokinetic studies in rats with truncated M-CSF forms.

Related Experiment Videos

  • Chemical modification of rM-CSF with polyethylene glycol (PEG).
  • X-ray crystallography of rM-CSF to determine its structural core.
  • Site-directed mutagenesis to identify receptor-binding residues.
  • Expression and purification of soluble M-CSF receptor (c-fms).
  • Biophysical analysis (size exclusion chromatography, light scattering) of M-CSF/receptor complexes.

    • Main Results:

    • Refolded nonglycosylated rM-CSF is functionally equivalent to glycosylated forms in vitro.
    • Truncated M-CSF forms exhibit increased circulating half-life with higher molecular weight.
    • PEGylation of rM-CSF significantly prolongs its in vivo half-life.
    • M-CSF possesses a four-helical-bundle structural core, with specific residues mediating receptor binding.
    • Mutagenesis confirmed the role of helix A and C residues in receptor interaction.
    • Soluble c-fms receptor binds to dimeric M-CSF, with each M-CSF binding two receptor molecules, supporting receptor dimerization in signaling.

    Conclusions:

    • Structural modifications can yield functionally active M-CSF variants with improved pharmacokinetic properties.
    • Understanding M-CSF's structure-function relationship and receptor interactions is key to its therapeutic potential.
    • M-CSF signaling involves receptor dimerization, mediated by specific structural features of M-CSF.