1KUV

X-ray Crystallographic Studies of Serotonin N-acetyltransferase Catalysis and Inhibition


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.00 Å
  • R-Value Free: 0.250 
  • R-Value Work: 0.203 
  • R-Value Observed: 0.226 

wwPDB Validation   3D Report Full Report


This is version 1.6 of the entry. See complete history


Literature

X-ray crystallographic studies of serotonin N-acetyltransferase catalysis and inhibition.

Wolf, E.De Angelis, J.Khalil, E.M.Cole, P.A.Burley, S.K.

(2002) J Mol Biol 317: 215-224

  • DOI: https://doi.org/10.1006/jmbi.2001.5371
  • Primary Citation of Related Structures:  
    1KUV, 1KUX, 1KUY

  • PubMed Abstract: 

    The structure of serotonin N-acetyltransferase (also known as arylalkylamine N-acetyltransferase; AANAT) bound to a potent bisubstrate analog inhibitor has been determined at 2.0 A resolution using a two-edge (Se, Br) multiwavelength anomalous diffraction (MAD) experiment. This acetyl-CoA dependent enzyme is a member of the GCN5-related family of N-acetyltransferases (GNATs), which share four conserved sequence motifs (A-D). In serotonin N-acetyltransferase, motif A adopts an alpha/beta conformation characteristic of the phylogenetically invariant cofactor binding site seen in all previously characterized GNATs. Motif B displays a significantly lower level of conservation among family members, giving rise to a novel alpha/beta structure for the serotonin binding slot. Utilization of a brominated CoA-S-acetyl-tryptamine-bisubstrate analog inhibitor and the MAD method permitted conclusive identification of two radically different conformations for the tryptamine moiety in the catalytic site (cis and trans). A second high-resolution X-ray structure of the enzyme bound to a bisubstrate analog inhibitor, with a longer tether between the acetyl-CoA and tryptamine moieties, demonstrates only the trans conformation. Given a previous proposal that AANAT can catalyze an alkyltransferase reaction in a conformationally altered active site relative to its acetyltransferase activity, it is possible that the two conformations of the bisubstrate analog observed crystallographically correspond to these alternative reaction pathways. Our findings may ultimately lead to the design of analogs with improved AANAT inhibitory properties for in vivo applications.


  • Organizational Affiliation

    Laboratories of Molecular Biophysics, The Rockefeller University, New York, NY 10021, USA.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Serotonin N-acetyltransferase207Ovis ariesMutation(s): 0 
Gene Names: U29663
EC: 2.3.1.87
UniProt
Find proteins for Q29495 (Ovis aries)
Explore Q29495 
Go to UniProtKB:  Q29495
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupQ29495
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 2 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
CA5
Query on CA5

Download Ideal Coordinates CCD File 
C [auth A]COA-S-ACETYL 5-BROMOTRYPTAMINE
C33 H47 Br N9 O17 P3 S
BBDVCGJBELWXIQ-GMHMEAMDSA-N
MG
Query on MG

Download Ideal Coordinates CCD File 
B [auth A]MAGNESIUM ION
Mg
JLVVSXFLKOJNIY-UHFFFAOYSA-N
Binding Affinity Annotations 
IDSourceBinding Affinity
CA5 Binding MOAD:  1KUV Ki: 22 (nM) from 1 assay(s)
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.00 Å
  • R-Value Free: 0.250 
  • R-Value Work: 0.203 
  • R-Value Observed: 0.226 
  • Space Group: C 2 2 21
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 53.313α = 90
b = 68.716β = 90
c = 89.725γ = 90
Software Package:
Software NamePurpose
SnBphasing
MLPHAREphasing
DMmodel building
CNSrefinement
MARMADdata reduction
DENZOdata reduction
SCALEPACKdata scaling
DMphasing

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2002-03-22
    Type: Initial release
  • Version 1.1: 2008-04-27
    Changes: Version format compliance
  • Version 1.2: 2011-07-13
    Changes: Version format compliance
  • Version 1.3: 2011-11-16
    Changes: Atomic model
  • Version 1.4: 2017-10-11
    Changes: Refinement description
  • Version 1.5: 2021-02-03
    Changes: Database references, Derived calculations, Structure summary
  • Version 1.6: 2024-02-14
    Changes: Data collection, Database references