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

Intersubunit disulfide-bonded lambda-Cro protein.

M Shirakawa1, H Matsuo, Y Kyogoku

  • 1Institute for Protein Research, Osaka University, Japan.

Protein Engineering
|June 1, 1991
PubMed
Summary

Mutating the Cro protein (Cro VC55) creates a stable dimer that is heat-stable but has reduced DNA-binding activity. This suggests subunit flexibility enhances DNA interaction affinity in wild Cro protein.

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

Isolation and identification of C-type natriuretic peptide in human monocytic cell line, THP-1.

Biochemical and biophysical research communications·1992
Same author

Stable expression of natriuretic peptide receptors: effects of HS-142-1, a non-peptide ANP antagonist.

Biochemical and biophysical research communications·1992
Same author

Gestational age-dependent dual action of epidermal growth factor on human placenta early in gestation.

The Journal of clinical endocrinology and metabolism·1992
Same author

[The prevalence and the clinical characteristics of silent myocardial ischemia detected by stress thallium scintigraphy].

Kaku igaku. The Japanese journal of nuclear medicine·1992
Same author

Inherited heterozygous protein C deficiency and dysfunctional protein S with recurrent venous thrombotic diseases: a study of three generations of a Japanese family.

Internal medicine (Tokyo, Japan)·1992
Same author

Cloning and characterization of an androgen-induced growth factor essential for the androgen-dependent growth of mouse mammary carcinoma cells.

Proceedings of the National Academy of Sciences of the United States of America·1992

Area of Science:

  • Molecular Biology
  • Protein Engineering
  • Biochemistry

Background:

  • The Cro protein of bacteriophage lambda is a key transcriptional regulator.
  • Understanding protein-dimerization and DNA-binding is crucial for gene regulation studies.

Purpose of the Study:

  • To investigate the role of the dimer interface in Cro protein function.
  • To engineer a more stable Cro protein variant and assess its DNA-binding properties.

Main Methods:

  • Site-directed mutagenesis to introduce cysteine at position 55 (Cro VC55).
  • Spectroscopic techniques including UV-Circular Dichroism (UV-CD) and Nuclear Magnetic Resonance (NMR).
  • Photochemically Induced Dynamic Nuclear Polarization (CIDNP) for conformational analysis.

Main Results:

  • Cro VC55 spontaneously formed a stable disulfide-bonded dimer.
  • The mutant protein exhibited enhanced heat stability while retaining wild-type conformation.
  • Specific DNA-binding activity was significantly reduced in Cro VC55.
  • Cro VC55 did not undergo conformational changes upon OR3 complex formation, unlike wild Cro.

Conclusions:

  • Disulfide-bonded dimerization of Cro VC55 leads to increased stability but decreased DNA affinity.
  • The flexibility or 'loosening' of wild Cro subunits upon DNA binding enhances affinity to operator DNA (OR3).
  • Induced fitting plays a critical role in the specific interaction between Cro and its operator DNA.

Related Experiment Videos