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

Recombinant human prorenin from CHO cells: expression and purification.

T F Holzman1, C C Chung, R Edalji

  • 1Protein Biochemistry, Pharmaceutical Discovery Research, Abbott Laboratories, Illinois 60064.

Journal of Protein Chemistry
|December 1, 1990
PubMed
Summary

Researchers successfully produced and isolated human prorenin in Chinese hamster ovary (CHO) cells. This recombinant protein, purified using chromatography, was characterized for its enzymatic activity, paving the way for further renin-related studies.

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

Designing e-consent protocols for pragmatic clinical trials: case studies from a UKCRC clinical trials unit.

Trials·2024
Same author

[Roles and mechanisms of m<sup>6</sup>A modification regulating <i>RP11-426A6.5</i> in laryngeal squamous cell carcinoma].

Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery·2023
Same author

Improving the electrostatic design of the Jefferson Lab 300 kV DC photogun.

The Review of scientific instruments·2022
Same author

Timing of procedural stroke and death in asymptomatic patients undergoing carotid endarterectomy: individual patient analysis from four RCTs.

The British journal of surgery·2020
Same author

Cells respond to deletion of CAV1 by increasing synthesis of extracellular matrix.

PloS one·2018
Same author

Mechanisms and mitigating factors for venous thromboembolism in chronic kidney disease: the REGARDS study.

Journal of thrombosis and haemostasis : JTH·2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Recombinant Protein Expression

Background:

  • Human prorenin is a precursor to renin, an enzyme critical in blood pressure regulation.
  • Efficient methods for producing and isolating prorenin are essential for studying the renin-angiotensin system.

Purpose of the Study:

  • To develop a system for the expression and secretion of human preprorenin in a mammalian cell line.
  • To establish a purification protocol for recombinant human prorenin.
  • To characterize the biochemical properties of the purified prorenin and its activated renin form.

Main Methods:

  • Construction of an expression vector containing human preprorenin cDNA and a dihydrofolate reductase (dhfr) selection marker.
  • Transfection of the vector into Chinese hamster ovary (CHO) DXB-11 cells.

Related Experiment Videos

  • Purification of secreted prorenin using filtration, concentration, dialysis, batch extraction, blue-dye chromatography, and size-exclusion chromatography.
  • Characterization of prorenin and activated renin using SDS-PAGE, N-terminal sequencing, sugar composition analysis, and enzyme activity assays.

    • Main Results:

    • High-level secretion of recombinant human prorenin (1-5 mg/L) into cell culture media by CHO cells.
    • Isolation of a single, purified prorenin protein band (Mr ~40,000) by preparative chromatography.
    • Characterization of the N-terminal sequence and glycosylation of prorenin, and the N-terminal sequence and pH-activity profile of trypsin-activated renin.

    Conclusions:

    • The study successfully established a method for producing and purifying biologically active human prorenin in a recombinant mammalian system.
    • The characterized recombinant prorenin and renin provide valuable tools for investigating the renin-angiotensin system and its role in physiological processes.