Allosteric control of three B12-dependent (class II) ribonucleotide reductases. Implications for the evolution of ribonucleotide reduction

• Published in: The Journal of biological chemistry 1999-03, Vol.274 (11), p.7182-7189
• Main Authors: Eliasson, R, Pontis, E, Jordan, A, Reichard, P
• Format: Article
• Language: English
• Subjects: Allosteric Regulation; Amino Acid Sequence; Biopolymers; Lactobacillus - enzymology; Medicin och hälsovetenskap; Molecular Sequence Data; Oxidation-Reduction; Protein Binding; Ribonucleotide Reductases - chemistry; Ribonucleotide Reductases - metabolism; Ribonucleotides - metabolism; Sequence Homology, Amino Acid; Thermoplasma - enzymology; Thermotoga maritima - enzymology; Vitamin B 12 - metabolism
• ISSN: 0021-9258
• DOI: 10.1074/jbc.274.11.7182

Three separate classes of ribonucleotide reductases are known, each with a distinct protein structure. One common feature of all enzymes is that a single protein generates each of the four deoxyribonucleotides. Class I and III enzymes contain an allosteric substrate specificity site capable of binding effectors (ATP or various deoxyribonucleoside triphosphates) that direct enzyme specificity. Some (but not all) enzymes contain a second allosteric site that binds only ATP or dATP. Binding of dATP to this site inhibits the activity of these enzymes. X-ray crystallography has localized the two sites within the structure of the Escherichia coli class I enzyme and identified effector-binding amino acids. Here, we have studied the regulation of three class II enzymes, one from the archaebacterium Thermoplasma acidophilum and two from eubacteria (Lactobacillus leichmannii and Thermotoga maritima). Each enzyme has an allosteric site that binds ATP or various deoxyribonucleoside triphosphates and that regulates its substrate specificity according to the same rules as for class I and III enzymes. dATP does not inhibit enzyme activity, suggesting the absence of a second active allosteric site. For the L. leichmannii and T. maritima enzymes, binding experiments also indicate the presence of only one allosteric site. Their primary sequences suggest that these enzymes lack the structural requirements for a second site. In contrast, the T. acidophilum enzyme binds dATP at two separate sites, and its sequence contains putative effector-binding amino acids for a second site. The presence of a second site without apparent physiological function leads to the hypothesis that a functional site was present early during the evolution of ribonucleotide reductases, but that its function was lost from the T. acidophilum enzyme. The other two B12 enzymes lost not only the function, but also the structural basis for the site. Also a large subgroup (Ib) of class I enzymes, but none of the investigated class III enzymes, has lost this site. This is further indirect evidence that class II and I enzymes may have arisen by divergent evolution from class III enzymes.