1CS4

COMPLEX OF GS-ALPHA WITH THE CATALYTIC DOMAINS OF MAMMALIAN ADENYLYL CYCLASE: COMPLEX WITH 2'-DEOXY-ADENOSINE 3'-MONOPHOSPHATE, PYROPHOSPHATE AND MG

Summary for 1CS4

• Entry DOI: 10.2210/pdb1cs4/pdb
• Related: 1cul
• Descriptor: TYPE V ADENYLATE CYCLASE, 5'-GUANOSINE-DIPHOSPHATE-MONOTHIOPHOSPHATE, TYPE II ADENYLATE CYCLASE, ... (11 entities in total)
• Functional Keywords: complex (lyase-hydrolase), hydrolase, signal transducing protein, cyclase, effector enzyme, lyase-hydrolase complex, lyase-lyase-signaling protein complex, lyase/lyase/signaling protein
• Biological source: Canis lupus familiaris (dog); More
• Cellular location: Membrane; Multi-pass membrane protein: P30803 P26769; Cell membrane; Lipid-anchor (By similarity): P04896
• Total number of polymer chains: 3
• Total formula weight: 96944.54

Authors

Tesmer, J.J.G.,Dessauer, C.A.,Sunahara, R.K.,Johnson, R.A.,Gilman, A.G.,Sprang, S.R. (deposition date: 1999-08-16, release date: 2001-01-10, Last modification date: 2024-02-07)

Primary citation

Tesmer, J.J.,Dessauer, C.W.,Sunahara, R.K.,Murray, L.D.,Johnson, R.A.,Gilman, A.G.,Sprang, S.R.
Molecular basis for P-site inhibition of adenylyl cyclase.
Biochemistry, 39:14464-14471, 2000

PubMed Abstract: P-site inhibitors are adenosine and adenine nucleotide analogues that inhibit adenylyl cyclase, the effector enzyme that catalyzes the synthesis of cyclic AMP from ATP. Some of these inhibitors may represent physiological regulators of adenylyl cyclase, and the most potent may ultimately serve as useful therapeutic agents. Described here are crystal structures of the catalytic core of adenylyl cyclase complexed with two such P-site inhibitors, 2'-deoxyadenosine 3'-monophosphate (2'-d-3'-AMP) and 2',5'-dideoxyadenosine 3'-triphosphate (2',5'-dd-3'-ATP). Both inhibitors bind in the active site yet exhibit non- or uncompetitive patterns of inhibition. While most P-site inhibitors require pyrophosphate (PP(i)) as a coinhibitor, 2',5'-dd-3'-ATP is a potent inhibitor by itself. The crystal structure reveals that this inhibitor exhibits two binding modes: one with the nucleoside moiety bound to the nucleoside binding pocket of the enzyme and the other with the beta and gamma phosphates bound to the pyrophosphate site of the 2'-d-3'-AMP.PP(i) complex. A single metal binding site is observed in the complex with 2'-d-3'-AMP, whereas two are observed in the complex with 2', 5'-dd-3'-ATP. Even though P-site inhibitors are typically 10 times more potent in the presence of Mn(2+), the electron density maps reveal no inherent preference of either metal site for Mn(2+) over Mg(2+). 2',5'-dd-3'-ATP binds to the catalytic core of adenylyl cyclase with a K(d) of 2.4 microM in the presence of Mg(2+) and 0.2 microM in the presence of Mn(2+). Pyrophosphate does not compete with 2',5'-dd-3'-ATP and enhances inhibition.

PubMed: 11087399
DOI: 10.1021/bi0015562

Experimental method

X-RAY DIFFRACTION (2.5 Å)