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

Studies on RNase T1 mutants affecting enzyme catalysis.

H P Grunert1, A Zouni, M Beineke

  • 1Institut für Kristallographie, Freie Universität Berlin, Federal Republic of Germany.

European Journal of Biochemistry
|April 10, 1991
PubMed
Summary

Mutating key amino acids in Aspergillus oryzae ribonuclease T1 (RNase T1) significantly reduced or eliminated its enzymatic activity. This research clarifies the catalytic mechanism of RNase T1, crucial for RNA processing.

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

Glucagon-like peptide-1 receptor agonists for treatment of diabetes and obesity: advantage of oral delivery.

Frontiers in drug delivery·2025
Same author

Detergent Choice Shapes the Solution Structures of Photosystems I and II: Implications for Crystallization and High-Resolution Studies.

The journal of physical chemistry. B·2025
Same author

Photoelectrochemistry of a photosystem I - Ferredoxin construct on ITO electrodes.

Bioelectrochemistry (Amsterdam, Netherlands)·2023
Same author

Current limits of structural biology: The transient interaction between cytochrome <i>c</i> <sub></sub> and photosystem I.

Current research in structural biology·2021
Same author

Practical implementation of the planar and spatial rotator in a complex tissue: the brain.

Journal of microscopy·2018
Same author

[Glaucoma in inpatient care in Germany in 2015].

Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft·2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Aspergillus oryzae ribonuclease T1 (RNase T1) is a vital enzyme for RNA processing.
  • Understanding the catalytic center of RNase T1 is key to elucidating its mechanism.
  • Site-directed mutagenesis is a powerful tool for probing enzyme function.

Purpose of the Study:

  • To investigate the role of specific amino acid residues in the catalytic center of RNase T1.
  • To characterize the enzymatic activity of RNase T1 mutants.
  • To gain further insights into the catalytic mechanism of RNase T1.

Main Methods:

  • Construction and characterization of site-directed mutants of RNase T1.
  • Expression of wild-type and mutant RNase T1 in an Escherichia coli overproducing strain.

Related Experiment Videos

  • Enzymatic activity assays (kcat/Km values) to quantify residual activity.

    • Main Results:

    • Mutants His92----Ala and His92----Phe exhibited complete loss of RNase T1 activity.
    • Mutants Glu58----Asp and Glu58----Gln retained 10% and 7% residual activity, respectively.
    • The His40----Thr mutant showed only 2% of wild-type RNase T1 activity.

    Conclusions:

    • Amino acid substitutions in the catalytic center significantly impact RNase T1 enzymatic activity.
    • The results support a mechanism involving Glu58, His40, and His92 in catalysis.
    • Structural and mechanistic insights into RNase T1 function were enhanced by these mutagenesis studies.