Crystal structure of calf spleen purine nucleoside phosphorylase with two full trimers in the asymmetric unit: important implications for the mechanism of catalysis

Journal of Molecular Biology
Agnieszka BzowskaJoachim Frank

Abstract

The crystal structure of the binary complex of trimeric purine nucleoside phosphorylase (PNP) from calf spleen with the acyclic nucleoside phosphonate inhibitor 2,6-diamino-(S)-9-[2-(phosphonomethoxy)propyl]purine ((S)-PMPDAP) is determined at 2.3A resolution in space group P2(1)2(1)2(1). Crystallization in this space group, which is observed for the first time with a calf spleen PNP crystal structure, is obtained in the presence of calcium atoms. In contrast to the previously described cubic space group P2(1)3, two independent trimers are observed in the asymmetric unit, hence possible differences between monomers forming the biologically active trimer could be detected, if present. Such differences would be expected due to third-of-the-sites binding documented for transition-state events and inhibitors. However, no differences are noted, and binding stoichiometry of three inhibitor molecules per enzyme trimer is observed in the crystal structure, and in the parallel solution studies using isothermal titration calorimetry and spectrofluorimetric titrations. Presence of phosphate was shown to modify binding stoichiometry of hypoxanthine. Therefore, the enzyme was also crystallized in space group P2(1)2(1)2(1) in the presence of...Continue Reading

References

Jan 1, 1978·Methods in Enzymology·J D StoecklerR E Parks
Mar 1, 1991·Acta Crystallographica. Section A, Foundations of Crystallography·T A JonesM Kjeldgaard
Jan 1, 1990·Zeitschrift Für Naturforschung. C, a Journal of Biosciences·A BzowskaD Shugar
May 1, 1993·Medicinal Research Reviews·J A Montgomery
Feb 1, 1996·Protein Science : a Publication of the Protein Society·E G Hutchinson, J M Thornton
Oct 8, 1997·Biochemistry·M D ErionS E Ealick
May 23, 1998·Advances in Experimental Medicine and Biology·E KulikowskaD Shugar
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Oct 25, 2000·Protein Science : a Publication of the Protein Society·F WangR H Angeletti
Jan 3, 2001·Acta Crystallographica. Section D, Biological Crystallography·M LuićA Bzowska
May 5, 2001·Pharmacology & Therapeutics·A BzowskaD Shugar
May 31, 2002·Acta Crystallographica. Section B, Structural Science·Frank H Allen, W D Samuel Motherwell
Jul 24, 2004·Acta Crystallographica. Section D, Biological Crystallography·Marija LuićAgnieszka Bzowska
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4

❮ Previous
Next ❯

Citations

Jan 12, 2010·Extremophiles : Life Under Extreme Conditions·Daniel F VisserDean Brady
Dec 17, 2009·Biochemical and Biophysical Research Communications·Katarzyna BreerAgnieszka Bzowska
Jul 1, 2008·Protein Expression and Purification·Katarzyna BreerAgnieszka Bzowska
Apr 26, 2005·Biophysical Chemistry·M DługoszJ M Antosiewicz
Sep 27, 2005·Journal of Molecular Biology·Humberto D'Muniz PereiraRichard Charles Garratt
Oct 7, 2015·Organic & Biomolecular Chemistry·Maëva ReverteMichael Smietana
Oct 13, 2005·Journal of Molecular Recognition : JMR·Abdessamad Ababou, John E Ladbury
Feb 14, 2008·Biochemistry·Mahmoud GhanemVern L Schramm
Jan 17, 2020·Journal of Chemical Information and Modeling·Maciej Pyrka, Maciej Maciejczyk
Oct 22, 2008·Archives of Biochemistry and Biophysics·Niya A Todorova, Frederick P Schwarz

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.