PDBsum entry 1cs4

Lyase/lyase/signaling protein

PDB id: 1cs4

Contents

Protein chains

189 a.a. *

190 a.a. *

329 a.a. *

Ligands

FOK

MES ×2

POP

101

GSP

Metals

_MG ×2

_CL

Waters ×77

* Residue conservation analysis

PDB id:

1cs4

Name:

Lyase/lyase/signaling protein

Title:

Complex of gs-alpha with the catalytic domains of mammalian adenylyl cyclase: complex with 2'-deoxy-adenosine 3'-monophosphate, pyrophosphate and mg

Structure:

Type v adenylate cyclase. Chain: a. Fragment: c1a domain. Synonym: atp pyrophosphate-lyase. Engineered: yes. Mutation: yes. Type ii adenylate cyclase. Chain: b. Fragment: c2a domain.

Source:

Canis lupus familiaris. Dog. Organism_taxid: 9615. Strain: familiaris. Organ: heart. Expressed in: escherichia coli. Expression_system_taxid: 562. Rattus norvegicus. Norway rat.

Biol. unit:

Trimer (from PQS)

Resolution:

2.50Å R-factor: 0.216 R-free: 0.264

Authors:

J.J.G.Tesmer,C.A.Dessauer,R.K.Sunahara,R.A.Johnson,A.G.Gilman, S.R.Sprang

Key ref:

J.J.Tesmer et al. (2000). Molecular basis for P-site inhibition of adenylyl cyclase. Biochemistry, 39, 14464-14471. PubMed id: 11087399 DOI: 10.1021/bi0015562

Date:

16-Aug-99 Release date: 10-Jan-01

Protein chain

P30803  (ADCY5_CANLF) -  Adenylate cyclase type 5 from Canis lupus familiaris

Seq: 1265 a.a.

Struc: 189 a.a.*

Protein chain

P26769  (ADCY2_RAT) -  Adenylate cyclase type 2 from Rattus norvegicus

Seq: 1090 a.a.

Struc: 190 a.a.

Protein chain

P04896  (GNAS2_BOVIN) -  Guanine nucleotide-binding protein G(s) subunit alpha isoforms short from Bos taurus

Seq: 394 a.a.

Struc: 329 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class 1: Chains A, B: E.C.4.6.1.1 - adenylate cyclase.
• Reaction: ATP = 3',5'-cyclic AMP + diphosphate

ATP (Bound ligand (Het Group name = GSP) matches with 90.91% similarity) = 3',5'-cyclic AMP + diphosphate (Bound ligand (Het Group name = POP) corresponds exactly)

• Cofactor: Pyridoxal 5'-phosphate

Pyridoxal 5'-phosphate (Bound ligand (Het Group name = 101) matches with 46.15% similarity)

• Enzyme class 2: Chain C: E.C.3.6.5.- - ?????

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1021/bi0015562

Biochemistry 39:14464-14471 (2000)

PubMed id: 11087399

Molecular basis for P-site inhibition of adenylyl cyclase.

J.J.Tesmer, C.W.Dessauer, R.K.Sunahara, L.D.Murray, R.A.Johnson, A.G.Gilman, S.R.Sprang.

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.