Related Experiment Video
Updated: Jul 23, 2025
12:49
Quantification of Site-specific Protein Lysine Acetylation and Succinylation Stoichiometry Using Data-independent Acquisition Mass Spectrometry
Published on: April 4, 2018
11.7K
Structural characterization of protein-material interfacial interactions using lysine reactivity profiling-mass
Zheyi Liu1,2, Shirui Yang1,2,3, Lingqiang Zhou1
1CAS Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Nature Protocols
|July 17, 2023
Summary
This study introduces a lysine reactivity profiling method using mass spectrometry to map protein-material interactions. This technique reveals critical binding sites and structural changes in protein-material hybrids.
Area of Science:
- Biomaterials Science
- Proteomics
- Analytical Chemistry
Background:
- Characterizing protein-material interactions is crucial for biomedical applications, including nanomaterial safety assessment and drug design.
- Understanding interfacial molecular details of protein-micro/nanomaterial hybrids is a significant challenge.
- Lysine residue accessibility and reactivity provide insights into protein behavior at interfaces.
Purpose of the Study:
- To develop and optimize a mass spectrometry-based strategy for profiling lysine reactivity in protein-material hybrids.
- To determine specific protein regions and binding sites involved in interactions with micro/nanomaterials.
- To provide a method for analyzing structural changes in proteins upon material binding.
Main Methods:
- Lysine reactivity profiling coupled with mass spectrometry.
- A two-step isotope dimethyl labeling strategy applied to protein-material hybrids under native and denaturing conditions.
- Optimization of protein digestion, elution, and data processing for interfacial structure analysis.
Main Results:
- The developed protocol accurately identifies protein-material interaction sites by quantifying changes in lysine residue reactivity.
- Comparative analysis of lysine reactivity under different conditions reveals native microenvironment influences.
- The method allows for the mapping of protein orientation, interaction regions, and modulated structures.
Conclusions:
- Lysine reactivity profiling-mass spectrometry is an effective strategy for characterizing protein-material interfaces.
- This method provides valuable data for the safety evaluation of biomedical nanomaterials and the design of novel nano-therapeutics.
- The protocol offers a comprehensive approach to analyzing complex protein-material interactions within a feasible timeframe (3-5 days).
