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Conformation of allantoicase in aqueous solution
E J 'S-Gravenmade1, C Van Der Drift, G D Vogels
1Department of Biophysical Chemistry and Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands.
Biochimica Et Biophysica Acta
|December 28, 1971
Summary
Researchers separated two forms of allantoicase, determining their molecular weights and structural properties. Lower helical content correlated with higher enzyme activity, suggesting similar structures despite size differences.
Area of Science:
- Biochemistry
- Enzymology
Background:
- Allantoicase is an enzyme involved in metabolic pathways.
- Understanding the structural and functional properties of different enzyme forms is crucial for biochemical research.
Purpose of the Study:
- To separate and characterize two distinct forms of allantoicase (0.9-S and 10.8-S).
- To investigate the molecular weight, structural conformation, and enzymatic activity of these allantoicase forms.
Main Methods:
- Separation using a 2H2O-H2O gradient.
- Analysis via sedimentation equilibrium, optical rotatory dispersion (ORD), circular dichroism (CD), and infrared spectroscopy.
- Determination of molecular weight, metal content, and amino acid composition.
Main Results:
- Two allantoicase forms, 0.9-S (MW ~1.1 x 10^4 g/mole) and 10.8-S (MW ~15.4 x 10^4 g/mole), were isolated.
- Both forms contain alpha-helix, beta conformation, and random coil structures.
- A Cotton effect at 418 nm indicated Mn2+ binding, with aromatic groups playing a role in substrate/inhibitor binding.
- Enzymes with lower alpha-helical content showed higher activity.
Conclusions:
- The 0.9-S and 10.8-S allantoicase forms exhibit similar physicochemical behavior and specific enzymatic activity.
- These findings suggest structural and conformational similarities between the two allantoicase forms, despite their size difference.