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

Toward efficient analysis of >70 kDa proteins with 100% sequence coverage.

A J Forbes1, M T Mazur, H M Patel

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School Boston, MA, USA.

Proteomics
|October 31, 2001
PubMed
Summary
This summary is machine-generated.

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

Green approach for synthesis of bioactive Hantzsch 1,4-dihydropyridine derivatives based on thiophene moiety via multicomponent reaction.

Royal Society open science·2017
Same author

Proteoforms in Peripheral Blood Mononuclear Cells as Novel Rejection Biomarkers in Liver Transplant Recipients.

American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons·2017
Same author

Assessment of water quality index of bore well water samples from some selected locations of South Gujarat, India.

Journal of environmental science & engineering·2015
Same author

Thiaminase I (42 kDa) heterogeneity, sequence refinement, and active site location from high-resolution tandem mass spectrometry.

Journal of the American Society for Mass Spectrometry·2013
Same author

Complete large-molecule high-resolution mass spectra from 50-femtomole microvolume injection.

Journal of the American Society for Mass Spectrometry·2013
Same author

Accurate base composition of double-strand DNA by mass spectrometry.

Journal of the American Society for Mass Spectrometry·2013
Same journal

Identification of Novel Interacting Proteins of FUZ and GPR161.

Proteomics·2026
Same journal

Light-Induced Proteomic Changes in Pseudomonas aeruginosa Biofilms.

Proteomics·2026
Same journal

Decade-Resolved Proteomic Profiling of Gastric Cancer FFPE Archives: Evaluating Storage-Associated Shifts and Signal Stability Over 50 Years.

Proteomics·2026
Same journal

Proteome-Scale Mining of Metal-Associated Proteins of Monkeypox Virus.

Proteomics·2026
Same journal

Optimized Sample Handling Minimizes Peptide Adsorption to Plastics to Enable High Sensitivity Evosep Based Chemical Proteomics.

Proteomics·2026
Same journal

Toward Predicting Pandemic Potential: A Comparative Analysis of Virus-Host Interactions Between Diverse Influenza A Viruses and the Human Innate Immune System.

Proteomics·2026
See all related articles

This study enhances protein mass spectrometry by using Lys-C enzyme digestion for efficient structural analysis of large proteins. This method improves peptide characterization, enabling more comprehensive protein structural determination.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Complete characterization of large proteins using mass spectrometry requires more efficient primary structural analysis.
  • Current methods face challenges with the size and complexity of intact large proteins.

Purpose of the Study:

  • To develop and illustrate a more efficient approach for the primary structural analysis of large proteins.
  • To assess the effectiveness of Lys-C digestion coupled with mass spectrometry for protein characterization.

Main Methods:

  • Proteins (159 kDa PchE, 199 kDa PchF) were digested using Lys-C enzyme.
  • Resulting peptides were analyzed using electrospray ionization and Fourier-transform mass spectrometry.
  • Isotopic resolution measurements were performed on a 4.7 Tesla instrument.

Related Experiment Videos

Main Results:

  • Lys-C digestion produced peptides ranging from 5 to 48 kDa, which are more amenable to mass spectrometry analysis.
  • For 199 kDa PchF (60% purity), 15% sequence coverage was achieved from 71 components.
  • Complete sequence coverage was obtained for 159 kDa PchE (>90% purity) using six Lys-C peptides.

Conclusions:

  • Lys-C digestion followed by mass spectrometry is an efficient strategy for large protein characterization.
  • This approach facilitates detailed primary structural analysis, even for complex protein mixtures.
  • The method shows promise for proteomic-scale protein characterization.