Nested Arg-specific bifunctional crosslinkers for MS-based structural analysis of proteins and protein assemblies
Qingrong Zhang1, Elizabeth Crosland, Daniele Fabris
1University of Maryland Baltimore County, Baltimore, MD 21228, United States.
Analytica Chimica Acta
|September 16, 2008
Summary
New bifunctional crosslinkers targeting Arg residues offer enhanced structural insights into biomolecules. These chemical probes, combined with mass spectrometry, reveal protein dynamics and aid in mapping complex molecular assemblies.
Area of Science:
- Biochemistry and Structural Biology
- Chemical Biology
- Molecular Biophysics
Background:
- Chemical probing and mass spectrometry are vital for biomolecular structure determination.
- Existing crosslinkers often target Cys and Lys residues, limiting structural coverage.
- Biomolecules with challenging size, solubility, or flexibility require advanced analytical tools.
Purpose of the Study:
- To develop novel Arg-specific bifunctional crosslinkers for enhanced biomolecular structural elucidation.
- To provide complementary structural information to existing Cys- and Lys-specific reagents.
- To investigate the utility of rigid, nested crosslinkers in mapping protein dynamics.
Main Methods:
- Synthesis of 1,4-phenyl-diglyoxal (PDG) and 4,4'-biphenyl-diglyoxal (BDG) Arg-specific crosslinkers.
- Electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry for analyzing crosslinking reactions.
- Proteolytic digestion and peptide mapping for identifying crosslinked sites in model proteins.
Main Results:
- PDG and BDG successfully formed mono- and bifunctional crosslinks with Arg residues in model proteins.
- Crosslinked sites correlated well with residue solvent accessibility and structural context.
- The rigid spacers of PDG and BDG provided insights into protein dynamics and flexibility.
Conclusions:
- Nested Arg-specific bifunctional crosslinkers are effective tools for biomolecular structure elucidation.
- These reagents offer complementary data to existing crosslinkers, improving structural coverage.
- The developed crosslinkers facilitate the study of flexible regions and aid in mapping larger biomolecular assemblies.
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