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 Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.5K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
6.5K
Signal Sequences and Sorting Receptors01:41

Signal Sequences and Sorting Receptors

13.5K
Signal sequences are short amino acid sequences that guide newly synthesized proteins to their proper location within the cell. Classical signal sequences are fifteen to sixty amino acids long and present at the N-terminus of a polypeptide chain. Each signal sequence has a conserved segment of basic residues towards their N terminus, a hydrophobic core, and a C-terminus rich in polar residues. The C-terminus also contains a signal cleavage site and features a -3 -1 sequence motif. The -3-1...
13.5K
DNA Topoisomerases02:02

DNA Topoisomerases

34.0K
Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
34.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Defined Nylon Oligomers Enable Mechanistic Insight Into Enzymatic Polyamide Depolymerization.

ChemSusChem·2026
Same author

Transcriptional insights into molecular events occurring during tuber maturation of tigernut (Cyperus esculentus L.).

Functional & integrative genomics·2026
Same author

The reliability and validity of rehabilitation set of the international classification of functioning, disability, and health in assessing Chinese tumor patients.

PloS one·2026
Same author

CECT Radiomics and Liver Fibrosis Markers for Predicting Early Recurrence in Older Patients with Hepatocellular Carcinoma.

Journal of hepatocellular carcinoma·2026
Same author

Genuine Directed Evolution In Test Tube (GENie).

bioRxiv : the preprint server for biology·2026
Same author

Simultaneous PET/fMRI distinguishes the separate and synergistic effects of T2DM and MCI on cerebral glucose metabolism and fALFF.

Scientific reports·2026
Same journal

Harnessing Naphthalimide Scaffolds for Sustainable CO<sub>2</sub> Utilization: A Metal-, Halide-, and Solvent-Free Photocatalytic CO<sub>2</sub> Cycloaddition via Sequential Two-Photon Activation.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same journal

Protein-Independent Liquid-Liquid Phase Separation of Adenosine Triphosphate Under Crowded Conditions.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same journal

A Unified Approach for the Synthesis of Conformationally Locked and sp<sup>2</sup>-sp<sup>3</sup> Fused Hybrids.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same journal

Decoding Heptazine Architectures: From Molecular Association to Structural Insight.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same journal

An Electrophilic Uridine Building Block for Post-Synthetic RNA Modification as Exemplified for Spin Labeling.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
Same journal

Recent Advances in Pd-Catalyzed Directed meta-C-H Olefination: Strategies and Outlook.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026
See all related articles

Related Experiment Video

Updated: Dec 8, 2025

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
08:34

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

Published on: February 5, 2020

7.0K

Enhancing Robustness of Sortase A by Loop Engineering and Backbone Cyclization.

Zhi Zou1,2, Diana M Mate2,3, Maximilian Nöth1,2

  • 1Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074, Aachen, Germany.

Chemistry (Weinheim an Der Bergstrasse, Germany)
|September 16, 2020
PubMed
Summary
This summary is machine-generated.

Researchers engineered a loop and cyclized the backbone of Staphylococcus aureus sortase A. This modified enzyme demonstrates improved stability for site-specific protein and peptide modifications.

More Related Videos

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.6K
Constructing Cyclic Peptides Using an On-Tether Sulfonium Center
07:11

Constructing Cyclic Peptides Using an On-Tether Sulfonium Center

Published on: September 28, 2022

3.0K

Related Experiment Videos

Last Updated: Dec 8, 2025

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
08:34

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

Published on: February 5, 2020

7.0K
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.6K
Constructing Cyclic Peptides Using an On-Tether Sulfonium Center
07:11

Constructing Cyclic Peptides Using an On-Tether Sulfonium Center

Published on: September 28, 2022

3.0K

Area of Science:

  • Biochemistry
  • Protein Engineering
  • Enzymology

Background:

  • Sortase A from Staphylococcus aureus is a transpeptidase enzyme crucial for bacterial cell wall anchoring.
  • Site-specific protein and peptide modification is essential for various biotechnological applications.
  • Improving the robustness of enzymes like Sortase A can enhance their utility in industrial processes.

Purpose of the Study:

  • To engineer a more robust variant of Staphylococcus aureus Sortase A.
  • To enhance the enzyme's stability for improved performance in site-specific protein and peptide modifications.
  • To investigate the structural modifications leading to enhanced enzyme robustness.

Main Methods:

  • Loop engineering and backbone cyclization of Staphylococcus aureus Sortase A.
  • Characterization of the modified enzyme's stability and activity.
  • Assays for site-specific protein and peptide modification using the engineered enzyme.

Main Results:

  • The loop-engineered and backbone-cyclized Sortase A exhibited significantly enhanced robustness.
  • The modified enzyme demonstrated improved stability under various conditions.
  • Successful application of the engineered enzyme in site-specific protein and peptide modifications with high efficiency.

Conclusions:

  • Loop engineering and backbone cyclization are effective strategies to enhance Sortase A robustness.
  • The engineered Sortase A variant offers improved performance for biotechnological applications.
  • This work provides a foundation for developing more stable enzymes for protein and peptide modification.