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Engineering surface loops of proteins--a preferred strategy for obtaining new enzyme function
A S el Hawrani1, K M Moreton, R B Sessions
1Molecular Recognition Centre, University of Bristol School of Medical Sciences, UK.
Trends in Biotechnology
|May 1, 1994
Summary
Enzyme redesign is feasible by altering surface amino acids without disrupting the protein structure. This review highlights successful modifications of Bacillus stearothermophilus lactate dehydrogenase (bsLDH) at solvent-exposed sites, particularly surface loops.
Area of Science:
- Biochemistry
- Protein Engineering
- Structural Biology
Background:
- Rational enzyme redesign requires preserving native protein structure during amino acid alteration for new functions.
- Combinatorial mutagenesis indicates that solvent-exposed protein surfaces tolerate random amino acid sequence variations best.
Purpose of the Study:
- To analyze successful redesigns of Bacillus stearothermophilus lactate dehydrogenase (bsLDH).
- To demonstrate that enzyme modifications can be achieved by targeting solvent-exposed residues.
Main Methods:
- Review of published studies on enzyme redesign.
- Analysis of residue variations in Bacillus stearothermophilus lactate dehydrogenase (bsLDH).
- Focus on modifications at solvent-exposed surfaces and surface loops.
Main Results:
- All analyzed residue variations in bsLDH were located at solvent-exposed surfaces.
- The majority of these variations occurred within surface loops connecting stable secondary structures.
- This strategy preserved the globular framework of the native protein.
Conclusions:
- Enzyme redesign is achievable by strategically altering solvent-exposed amino acids, particularly within surface loops.
- Targeting surface loops allows for functional modifications while maintaining protein structural integrity.
- This approach facilitates the development of novel enzyme functions without compromising the native protein framework.