Characterization of a novel mammalian phosphatase having sequence similarity to Schizosaccharomyces pombe PHO2 and Saccharomyces cerevisiae PHO13

Biochemistry
MacKevin I NdubuisilC M Crews

Abstract

p34, a specific p-nitrophenyl phosphatase (pNPPase) was identified and purified from the murine cell line EL4 in a screen for the intracellular molecular targets of the antiinflammatory natural product parthenolide. A BLAST search analysis revealed that it has a high degree of sequence similarity to two yeast alkaline phosphatases. We have cloned, sequenced, and expressed p34 as a GST-tagged fusion protein in Escherichia coli and an EE-epitope-tagged fusion protein in mammalian cells. Using p-nitrophenyl phosphate (pNPP) as a substrate, p34 is optimally active at pH 7.6 with a K(m) of 1.36 mM and K(cat) of 0.052 min(-1). Addition of 1 mM Mg(2+) to the reaction mixture increases its activity by 14-fold. Other divalent metal ions such as Co(2+) and Mn(2+) also stimulated the activity of the enzyme, while Zn(2+), Fe(2+), and Cu(2+) had no effect. Furthermore, both NaCl and KCl enhanced the activity of the enzyme, having maximal effect at 50 and 75 mM, respectively. The enzyme is inhibited by sodium orthovanadate but not by sodium fluoride or okadaic acid. Mutational analysis data suggest that p34 belongs to the group of phosphatases characterized by the sequence motif DXDX(T/V).

References

Jan 1, 1979·Annual Review of Biochemistry·E G Krebs, J A Beavo
Mar 1, 1992·Trends in Biochemical Sciences·J B VincentB A Averill
Jun 1, 1991·European Journal of Biochemistry·J W YangM E Schweingruber
Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J F BazanS J Pilkis
Jul 1, 1989·Annals of the Rheumatic Diseases·M PattrickM Doherty
Jan 1, 1986·The International Journal of Biochemistry·J W Sparks, D L Brautigan
Oct 10, 1973·Biochimica Et Biophysica Acta·J Attias, H Durand
May 12, 1972·Biochimica Et Biophysica Acta·J Attias, J L Bonnet
Feb 1, 1997·Protein Science : a Publication of the Protein Society·J Stukey, G M Carman
Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·K ShahK M Shokat
Jun 10, 1997·Proceedings of the National Academy of Sciences of the United States of America·N SinC M Crews
May 23, 1998·Annual Review of Immunology·S GhoshE B Kopp
Jun 11, 1998·The Journal of Biological Chemistry·J F ColletEmile Van Schaftingen
Oct 24, 1998·Bioorganic & Medicinal Chemistry·N SinC M Crews
Sep 1, 1999·Proceedings of the National Academy of Sciences of the United States of America·L MengC M Crews
Nov 24, 1999·The Journal of Biological Chemistry·J F ColletEmile Van Schaftingen
Feb 19, 2000·Current Opinion in Chemical Biology·C M Crews, R Mohan
Jul 25, 2000·The Cochrane Database of Systematic Reviews·M H PittlerE Ernst
Aug 15, 2000·Annual Review of Biophysics and Biomolecular Structure·A BishopK M Shokat

Citations

Jul 16, 2011·The Journal of Biological Chemistry·Hans C HürlimannBenoît Pinson
Apr 12, 2005·Annual Review of Pharmacology and Toxicology·Christophe Morisseau, Bruce D Hammock
Feb 8, 2003·Proceedings of the National Academy of Sciences of the United States of America·John W NewmanBruce D Hammock
Nov 16, 2004·Biochemical and Biophysical Research Communications·Wei Wang, Peter J Roach
Jun 14, 2016·Nature Chemical Biology·François CollardGuido T Bommer
Oct 31, 2006·Oncogene·T D Gilmore, M Herscovitch
Feb 1, 2008·The Journal of Biological Chemistry·Venky Sreedhar ReddyRam Rajasekharan

Related Concepts

Parthenolide, (1aR-(1aR*,4E,7aS*,10aS*,10bR*))-isomer
PHO2 protein, S cerevisiae
Metazoa
Analgesics, Anti-Inflammatory
Medebiotin
Cations, Divalent
Cations, Monovalent
Enzyme Activation
Enzyme Inhibitors
Yeast Proteins

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.

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.

Lipidomics & Rhinovirus Infection

Lipidomics can be used to examine the lipid species involved with pathogenic conditions, such as viral associated inflammation. Discovered the latest research on Lipidomics & Rhinovirus Infection.

Alzheimer's Disease: MS4A

Variants within the membrane-spanning 4-domains subfamily A (MS4A) gene cluster have recently been implicated in Alzheimer's disease in genome-wide association studies. Here is the latest research on Alzheimer's disease and MS4A.

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.

Torsion Dystonia

Torsion dystonia is a movement disorder characterized by loss of control of voluntary movements appearing as sustained muscle contractions and/or abnormal postures. Here is the latest research.

Generating Insulin-Secreting Cells

Reprogramming cells or using induced pluripotent stem cells to generate insulin-secreting cells has significant therapeutic implications for diabetics. Here is the latest research on generation of insulin-secreting cells.

Central Pontine Myelinolysis

Central Pontine Myelinolysis is a neurologic disorder caused most frequently by rapid correction of hyponatremia and is characterized by demyelination that affects the central portion of the base of the pons. Here is the latest research on this disease.

Epigenome Editing

Epigenome editing is the directed modification of epigenetic marks on chromatin at specified loci. This tool has many applications in research as well as in the clinic. Find the latest research on epigenome editing here.