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

Isolation of eukaryotic initiation factor 2 from rat brain.

C Calés, M Salinas, J L Fando

    Journal of Neurochemistry
    |October 1, 1985
    PubMed
    Summary

    Researchers purified eukaryotic initiation factor 2 (eIF-2) from rat brain, finding its activity decreased due to the loss of modulating factors during purification.

    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

    Advancing cardiovascular risk assessment: Real-time SCORES2 calculation through CDSS in primary care patients.

    Clinical biochemistry·2025
    Same author

    Quality of life in idiopathic pulmonary fibrosis in Latin American countries.

    BMC pulmonary medicine·2025
    Same author

    A PTHrP Gradient Drives Mandibular Condylar Chondrogenesis via Runx2.

    Journal of dental research·2023
    Same author

    Regulatory effects of miR-19a on MAD2 expression and tumorigenesis in gastric cancer.

    Genes & diseases·2023
    Same author

    Pilot study of women's perspectives when abnormal uterine bleeding occurs during perimenopause.

    Climacteric : the journal of the International Menopause Society·2022
    Same author

    What outcomes should researchers select, collect and report in pre-eclampsia research? A qualitative study exploring the views of women with lived experience of pre-eclampsia.

    BJOG : an international journal of obstetrics and gynaecology·2019

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Neuroscience

    Background:

    • Protein synthesis is crucial for brain development and function.
    • Eukaryotic initiation factor 2 (eIF-2) plays a key role in regulating protein synthesis initiation.
    • High rates of protein synthesis in developing rat brains suggest the presence of active translational machinery.

    Purpose of the Study:

    • To isolate and characterize eukaryotic initiation factor 2 (eIF-2) from the highly synthetic 4-day-old rat brain.
    • To investigate the composition and activity of purified eIF-2.
    • To understand factors influencing eIF-2 activity in the context of high protein synthesis rates.

    Main Methods:

    • Isolation of eIF-2 from salt-washed rat brain microsomes.
    • A three-step purification process involving heparin-Sepharose, phosphocellulose, and DEAE-cellulose chromatography.
    • Analysis of purified eIF-2 using SDS-PAGE to determine polypeptide composition and molecular weights.
    • Assay of eIF-2 activity through ternary complex formation with radiolabeled methionyl-tRNAi and GTP.

    Main Results:

    • Purified eIF-2 consisted of three polypeptides with molecular weights of 43,000, 54,000, and 59,000, achieving 90% purity.
    • The purified rat brain eIF-2 demonstrated the ability to form ternary complexes with [3H]methionyl-tRNAi and GTP.
    • Specific activity of eIF-2 decreased during purification, not correlating with electrophoretic purity.
    • An apparent loss of additional modulating factors during purification was observed, explaining the high activity in crude fractions.

    Conclusions:

    • Eukaryotic initiation factor 2 (eIF-2) was successfully isolated from developing rat brain tissue.
    • The purification process led to a loss of essential co-factors, resulting in reduced specific activity compared to crude extracts.
    • This highlights the importance of accessory factors in modulating eIF-2 function and overall protein synthesis rates in the brain.

    Related Experiment Videos