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

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

12.7K
We present a protocol to accurately quantitate proteins with isobaric labelling, extensive fractionation, bioinformatics tools, and quality control steps in combination with liquid chromatography interfaced to a high-resolution mass...
12.7K
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics11:40

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

2.9K
A neuronal lysosome proximity labeling proteomics protocol is described here to characterize the dynamic lysosomal microenvironment in human induced pluripotent stem cell-derived neurons. Lysosomal membrane proteins and proteins that interact with lysosomes (stably or transiently) can be accurately quantified in this method with excellent intracellular spatial resolution in live human...
2.9K
"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome06:31

"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome

3.1K
Here, we describe a proteomics workflow for characterization of the cell surface proteome of various cell types. This workflow includes cell surface protein enrichment, subsequent sample preparation, analysis using an LC-MS/MS platform, and data processing with specialized...
3.1K
Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

16.8K
Stable isotope labeling of peptides by reductive dimethylation (ReDi labeling) is a rapid, inexpensive strategy for accurate mass spectrometry-based quantitative proteomics. Here we demonstrate a robust method for preparation and analysis of protein mixtures using the ReDi approach that can be applied to nearly any sample type.
16.8K
Antibody Profiling by Luciferase Immunoprecipitation Systems (LIPS)12:19

Antibody Profiling by Luciferase Immunoprecipitation Systems (LIPS)

27.7K
The technical aspects of performing LIPS (Luciferase Immunoprecipitation Systems) are described. The overall approach involves expressing chimeric genes encoding antigens fused to Renilla luciferase (Ruc) in mammalian cells. Crude Ruc-antigen extracts are then prepared and, without purification, employed in immunoprecipitation assays to quantify...
27.7K
Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry10:36

Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry

6.0K
The experimental procedures for performing diethylpyrocarbonate-based covalent labeling with mass spectrometric detection are described. Diethylpyrocarbonate is simply mixed with the protein or protein complex of interest, leading to the modification of solvent accessible amino acid residues. The modified residues can be identified after proteolytic digestion and liquid chromatography/mass spectrometry...
6.0K

You might also read

Related Articles

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

Sort by
Same author

De-escalation of breast surgery for early stage breast cancer.

Japanese journal of clinical oncology·2026
Same author

Clinical implications of discordance in HER2 reassessment from HercepTest to 4B5 in metastatic breast cancer.

Breast cancer research and treatment·2026
Same author

Prognostic impact of progesterone receptor status in patients with breast cancer and isolated locoregional recurrence.

Breast cancer (Tokyo, Japan)·2026
Same author

Safety and Effectiveness of an Integrative Treatment of Acupuncture-Based Intervention in Survivors of Breast Cancer With Postmastectomy Pain Syndrome: Protocol for a Single-Center, Single-Arm Exploratory Trial.

JMIR research protocols·2026
Same author

Comprehensive High-Depth Proteomic Analysis of Plasma Extracellular Vesicle-Containing Preparations in CDKL5 Deficiency Disorder.

Biomedicines·2026
Same author

Decreased expression of Ly-1 antibody reactive clone (Lyar) triggers enhanced adipogenesis of bone marrow mesenchymal stromal cells in aged bone marrow.

PloS one·2026
Same journal

NMR Spectroscopy: Molecular Insights into Cell Wall Collapse and Oxidative Stress of <i>Escherichia coli</i> Induced by Imidazole-Activated Eutectic Solvents.

ACS omega·2026
Same journal

Enhanced Arsenite Remediation in Synthetic FeS<sub>2</sub>/Fe(II)-Containing Arsenic Wastewater via Epigallocatechin Gallate-Initiated Persulfate Activation.

ACS omega·2026
Same journal

Defect and Particle-Size Engineering as Mechanistic Drivers for Dye Uptake in a Zirconium Metal-Organic Framework.

ACS omega·2026
Same journal

Biogeochemical Assessment of Short-Term Hydrogen Storage in Methane Reservoirs with Field Sample Characterization and Reactor Experiments.

ACS omega·2026
Same journal

Combined Effects of Halloysite Nanotubes, Nucleating Agent, and Thermal Annealing on the Printability and Mechanical Performances of 3D-Printable Polypropylene Random Copolymer-Based Composites.

ACS omega·2026
Same journal

Effect of MoS<sub>2</sub> Interfacial Engineering across MAPbI<sub>3</sub>, FAPbI<sub>3</sub>, and CsPbI<sub>3</sub> Perovskite Solar Cells.

ACS omega·2026
See all related articles

Related Experiment Video

Updated: Jan 20, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

12.7K

Rapid and Label-Free Structural Proteomics Using One-Step Swift Trypsin LiP-MS.

Yasuomi Miyashita1,2,3, Ryo Konno1, Satoshi Ogasawara3,4

  • 1Department of Applied Genomics, Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan.

ACS Omega
|January 19, 2026
PubMed
Summary
This summary is machine-generated.

A new Swift Trypsin Limited Proteolysis Mass Spectrometry (STLiP-MS) method simplifies proteomic analysis. This rapid technique accurately detects protein structural changes and predicts interaction sites, enhancing drug target identification.

More Related Videos

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
11:40

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Published on: June 23, 2022

2.9K
"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome
06:31

"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome

Published on: March 24, 2023

3.1K

Related Experiment Videos

Last Updated: Jan 20, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

12.7K
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
11:40

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Published on: June 23, 2022

2.9K
"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome
06:31

"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome

Published on: March 24, 2023

3.1K

Area of Science:

  • Proteomics
  • Structural Biology
  • Biochemistry

Background:

  • Limited proteolysis mass spectrometry (LiP-MS) is crucial for studying protein conformational changes.
  • Conventional LiP-MS workflows are complex, reducing reproducibility and sensitivity.
  • A simplified, rapid, and sensitive method is needed for proteome-wide structural analysis.

Purpose of the Study:

  • To develop a simplified one-step protocol for LiP-MS, named Swift Trypsin LiP-MS (STLiP-MS).
  • To enhance the sensitivity and reproducibility of proteomic structural analysis.
  • To demonstrate the method's applicability in identifying protein-protein interactions and drug targets.

Main Methods:

  • Developed STLiP-MS using a trypsin-immobilized spin column and high-speed centrifugation for surface-limited proteolysis.
  • Optimized the protocol by suppressing predigestion and enabling immediate enzyme inactivation.
  • Applied STLiP-MS to HEK293 cell extracts and the adenosine A2A receptor (A2A-BRIL).

Main Results:

  • STLiP-MS identified 286 proteins with conformational changes in HEK293 cells, with 37 linked to phosphatase activity.
  • Enhanced sensitivity detected 799 proteins with structural alterations, 77 enriched in phosphatase-related categories.
  • Validated antibody-induced protection on A2A-BRIL and confirmed interaction sites using cryogenic electron microscopy.

Conclusions:

  • STLiP-MS is a rapid, robust, and sensitive platform for label-free detection of local structural changes under near-physiological conditions.
  • The method accurately predicts protein-protein interaction sites.
  • STLiP-MS shows significant promise for structural proteomics and drug target identification.