Bicarbonate activation of adenylyl cyclase via promotion of catalytic active site closure and metal recruitment

Nat Struct Mol Biol. 2005 Jan;12(1):32-7. doi: 10.1038/nsmb880. Epub 2004 Dec 26.

Abstract

In an evolutionarily conserved signaling pathway, 'soluble' adenylyl cyclases (sACs) synthesize the ubiquitous second messenger cyclic adenosine 3',5'-monophosphate (cAMP) in response to bicarbonate and calcium signals. Here, we present crystal structures of a cyanobacterial sAC enzyme in complex with ATP analogs, calcium and bicarbonate, which represent distinct catalytic states of the enzyme. The structures reveal that calcium occupies the first ion-binding site and directly mediates nucleotide binding. The single ion-occupied, nucleotide-bound state defines a novel, open adenylyl cyclase state. In contrast, bicarbonate increases the catalytic rate by inducing marked active site closure and recruiting a second, catalytic ion. The phosphates of the bound substrate analogs are rearranged, which would facilitate product formation and release. The mechanisms of calcium and bicarbonate sensing define a reaction pathway involving active site closure and metal recruitment that may be universal for class III cyclases.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / pharmacology
  • Adenylyl Cyclases / chemistry*
  • Adenylyl Cyclases / metabolism*
  • Amino Acid Sequence
  • Animals
  • Bicarbonates / chemistry
  • Bicarbonates / pharmacology*
  • Binding Sites / drug effects
  • Calcium / metabolism*
  • Catalytic Domain / drug effects*
  • Crystallography, X-Ray
  • Cyanobacteria / enzymology
  • Cyanobacteria / genetics
  • Enzyme Activation / drug effects
  • Humans
  • Magnesium / metabolism*
  • Mammals
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Thionucleotides / metabolism

Substances

  • Bicarbonates
  • Thionucleotides
  • adenosine 5'-(1-thio)triphosphate
  • Adenosine Triphosphate
  • Adenylyl Cyclases
  • Magnesium
  • alpha,beta-methyleneadenosine 5'-triphosphate
  • Calcium

Associated data

  • PDB/1AZS
  • PDB/1WC0
  • PDB/1WC1
  • PDB/1WC3
  • PDB/1WC4
  • PDB/1WC5
  • PDB/1WC6