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

Protein engineering studies on C1r and C1s

P Závodszky1, P Gál, S Cseh

  • 1Institute of Enzymology, Hungarian Academy of Sciences, Budapest.

Behring Institute Mitteilungen
|December 1, 1993
PubMed
Summary

Recombinant C1r and C1s proteins were successfully expressed in insect cells, revealing crucial roles for specific domains in complement system component C1 complex formation and autoactivation. These findings advance understanding of C1 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

Safety of a gastropexy device in infants and young children in percutaneous endoscopic gastrostomy tube placement.

Scientific reports·2025
Same author

Cardiovascular risk factors: The effects of ageing and smoking on the immune system, an observational clinical study.

Frontiers in immunology·2022
Same author

Effect of retinol in the vitrification medium on viability of vitrified ovine preantral follicles and expression of key developmental and apoptosis related genes.

Cryo letters·2022
Same author

Effect of different vitrification protocols on post thaw viability and gene expression of ovine preantral follicles.

Theriogenology·2021
Same author

Genistein does not inhibit TGF-beta1-induced conversion of human dermal fibroblasts to myofibroblasts.

Physiological research·2021
Same author

Effect of retinol as antioxidant on the post-thaw viability and the expression of apoptosis and developmental competence-related genes of vitrified preantral follicles in buffalo (Bubalus bubalis).

Reproduction in domestic animals = Zuchthygiene·2021

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • The complement system is a critical part of innate immunity.
  • Component C1, the first component of the classical complement pathway, comprises C1q, C1r, and C1s.
  • Understanding the structure-function relationships of C1r and C1s is essential for elucidating complement activation.

Purpose of the Study:

  • To express biologically active recombinant C1r and C1s proteins in insect cells.
  • To investigate the roles of specific domains within C1r and C1s in complex formation and autoactivation.
  • To analyze the impact of post-translational modifications on C1r and C1s function.

Main Methods:

  • Recombinant expression of C1r and C1s using Autographa californica nuclear polyhedrosis virus in insect cells.
  • Hemolytic assays to assess biological activity of recombinant proteins.
  • Construction and expression of deletion and point mutants of C1r cDNA.
  • Construction and expression of hybrid C1r-C1s cDNA molecules.

Main Results:

  • High yield of biologically active recombinant C1r and C1s proteins obtained.
  • Recombinant C1r and C1s exhibited biological activity, though with differences in glycosylation compared to serum proteins.
  • Domains I and II of C1r are essential for tetramer formation and regulate autoactivation.
  • A hybrid C1s construct demonstrated modulated function influenced by the C1r portion.
  • Point mutations in C1r catalytic domain affected its autoactivation rate and zymogen stability.

Conclusions:

  • Insect cells can produce functional recombinant C1r and C1s.
  • Specific domains of C1r play critical roles in C1 complex assembly and regulation.
  • Post-translational modifications in insect cells, while different, do not impede essential functions.
  • Mutational analysis provides insights into the intramolecular regulation of C1 autoactivation.

Related Experiment Videos