Cloning, expression, characterisation and three-dimensional structure determination ofCaenorhabditis elegansspermidine synthase
Open Access
- 5 October 2005
- journal article
- Published by Wiley in FEBS Letters
- Vol. 579 (27), 6037-6043
- https://doi.org/10.1016/j.febslet.2005.09.050
Abstract
The polyamine synthesis enzyme spermidine synthase (SPDS) has been cloned from the model nematode Caenorhabditis elegans. Biochemical characterisation of the recombinantly expressed protein revealed a high degree of similarity to other eukaryotic SPDS with the exception of a low affinity towards the substrate decarboxylated S‐adenosylmethionine (K m = 110 μM) and a less pronounced feedback inhibition by the second reaction product 5’‐methylthioadenosine (IC50 = 430 μM). The C. elegans protein that carries a nematode‐specific insertion of 27 amino acids close to its N‐terminus was crystallized, leading to the first X‐ray structure of a dimeric eukaryotic SPDS.Keywords
This publication has 30 references indexed in Scilit:
- The crystal structure of spermidine synthase with a multisubstrate adduct inhibitorNature Structural & Molecular Biology, 2001
- Refinement of Macromolecular Structures by the Maximum-Likelihood MethodActa Crystallographica Section D-Biological Crystallography, 1997
- Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constantsJournal of Applied Crystallography, 1993
- PROCHECK: a program to check the stereochemical quality of protein structuresJournal of Applied Crystallography, 1993
- Improved methods for building protein models in electron density maps and the location of errors in these modelsActa Crystallographica Section A Foundations of Crystallography, 1991
- Human Spermidine Synthase: Cloning and Primary StructureDNA and Cell Biology, 1990
- POLYAMINESAnnual Review of Biochemistry, 1984
- Role of Polyamines in the Control of Cell Proliferation and DifferentiationDifferentiation, 1981
- On the treatment of negative intensity observationsActa Crystallographica Section A, 1978
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976