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Solution structure of an isolated antibody VL domain
K L Constantine1, M S Friedrichs, W J Metzler
1Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543.
Journal of Molecular Biology
|February 11, 1994
Summary
The solution structure of the anti-digoxin antibody 26-10 VL domain was determined, revealing it weakly associates and rapidly forms monomers/dimers. This study provides the first structure of an antibody VL domain without fixed quaternary interactions.
Area of Science:
- Structural Biology
- Biochemistry
- Immunology
Background:
- Antibody variable light (VL) domains are crucial for antigen binding.
- Understanding VL domain structure in isolation is key to deciphering antigen-antibody interactions.
- Previous structural studies often involved VL domains within a fixed quaternary context.
Purpose of the Study:
- To determine the solution structure of the isolated VL domain of the anti-digoxin antibody 26-10.
- To investigate the quaternary interactions and conformational flexibility of the VL domain.
- To provide insights into antibody structure-function relationships.
Main Methods:
- Heteronuclear multi-dimensional nuclear magnetic resonance (n.m.r.) spectroscopy.
- Analytical ultracentrifugation to assess dimerization.
- Structure calculation using DIANA and refinement with X-PLOR.
Main Results:
- The VL domain exhibits weak association (Kd = 2.5 mM) and rapid monomer/dimer equilibrium.
- A detailed solution structure ensemble was generated using extensive n.m.r. constraints.
- Comparison with X-ray structure reveals minor differences, primarily in CDR loops and beta-strands.
Conclusions:
- The 26-10 VL domain conformation is mainly governed by intradomain interactions.
- Quaternary VL-VH association induces only minor conformational adjustments.
- This work presents the first structure of an antibody VL domain determined in the absence of fixed quaternary interactions.