The structure of adenylosuccinate lyase, an enzyme with dual activity in the de novo purine biosynthetic pathway

• Published in: Structure (London) 2000-02, Vol.8 (2), p.163-174
• Main Authors: Toth, Eric A, Yeates, Todd O
• Format: Article
• Language: English
• Subjects: Adenylosuccinate Lyase - chemistry; Adenylosuccinate Lyase - genetics; Adenylosuccinate Lyase - metabolism; Amino Acid Sequence; Base Sequence; Binding Sites; Crystallography, X-Ray; DNA Primers; Humans; Lyase; Models, Molecular; Molecular Sequence Data; Mutation; Protein Conformation; Purine biosynthesis; Purines - biosynthesis; Sequence Homology, Amino Acid; Thermotoga maritima - enzymology; Thermotoga maritima - metabolism; β-elimination superfamily
• ISSN: 0969-2126; 1878-4186
• DOI: 10.1016/S0969-2126(00)00092-7

Background: Adenylosuccinate lyase is an enzyme that plays a critical role in both cellular replication and metabolism via its action in the de novo purine biosynthetic pathway. Adenylosuccinate lyase is the only enzyme in this pathway to catalyze two separate reactions, enabling it to participate in the addition of a nitrogen at two different positions in adenosine monophosphate. Both reactions catalyzed by adenylosuccinate lyase involve the β-elimination of fumarate. Enzymes that catalyze this type of reaction belong to a superfamily, the members of which are homotetramers. Because adenylosuccinate lyase plays an integral part in maintaining proper cellular metabolism, mutations in the human enzyme can have severe clinical consequences, including mental retardation with autistic features. Results: The 1.8 Å crystal structure of adenylosuccinate lyase from Thermotoga maritima has been determined by multiwavelength anomalous dispersion using the selenomethionine-substituted enzyme. The fold of the monomer is reminiscent of other members of the β-elimination superfamily. However, its active tetrameric form exhibits striking differences in active-site architecture and cleft size. Conclusions: This first structure of an adenylosuccinate lyase reveals that, along with the catalytic base (His141) and the catalytic acid (His68), Gln212 and Asn270 might play a vital role in catalysis by properly orienting the succinyl moiety of the substrates. We propose a model for the dual activity of adenylosuccinate lyase: a single 180° bond rotation must occur in the substrate between the first and second enzymatic reactions. Modeling of the pathogenic human S413P mutation indicates that the mutation destabilizes the enzyme by disrupting the C-terminal extension.