Journal
/
Biochemistry
/
OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

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

DOI:
10.3791/60774-v

•

08:34 min

•

February 05, 2020

•

, , , , , ,
1State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering,Nanjing University

Chapters

  • 00:05Introduction
  • 00:43Functionalization of Coverslip and Cantilever Surface
  • 04:31Stepwise Polyprotein Preparation with Controlled Sequences
  • 05:44AFM Experiment Measurement and Data Analysis
  • 06:54Results: AFM-based SMFS Measurements, UV-Vis Absorbance Spectra and SDS-PAGE Gel Results
  • 07:51Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to conjugate protein monomer by enzymes forming protein polymer with a controlled sequence and immobilize it on the surface for single-molecule force spectroscopy studies.

Tags

OaAEP1Enzymatic SynthesisPolymerized ProteinSingle-molecule Force SpectroscopyProtein OligomerPolymerization DegreeProtein-based MaterialCysteinePotassium ChromateSulfuric AcidAPTESSulfo-SMCCGL-ELP-50 Cys-proteinCantilever Surface

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/60774?article_type=v

Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues
check_url/60774?article_type=v

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
check_url/60774?article_type=v

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
check_url/60774?article_type=v

Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers
check_url/60774?article_type=v

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions
check_url/60774?article_type=v

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
check_url/60774?article_type=v

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
check_url/60774?article_type=v

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
check_url/60774?article_type=v

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
check_url/60774?article_type=v

Single Molecule Fluorescence Energy Transfer Study of Ribosome Protein Synthesis

Read Article