The phosphate site of trehalose phosphorylase from Schizophyllum commune probed by site-directed mutagenesis and chemical rescue studies.

The FEBS Journal
Christiane Goedl, Bernd Nidetzky

Abstract

Schizophyllum communealpha,alpha-trehalose phosphorylase utilizes a glycosyltransferase-like catalytic mechanism to convert its disaccharide substrate into alpha-d-glucose 1-phosphate and alpha-d-glucose. Recruitment of phosphate by the free enzyme induces alpha,alpha-trehalose binding recognition and promotes the catalytic steps. Like the structurally related glycogen phosphorylase and other retaining glycosyltransferases of fold family GT-B, the trehalose phosphorylase contains an Arg507-XXXX-Lys512 consensus motif (where X is any amino acid) comprising key residues of its putative phosphate-binding sub-site. Loss of wild-type catalytic efficiency for reaction with phosphate (kcat/Km=21,000 m(-1).s(-1)) was dramatic (>or=10(7)-fold) in purified Arg507-->Ala (R507A) and Lys512-->Ala (K512A) enzymes, reflecting a corresponding change of comparable magnitude in kcat (Arg507) and Km (Lys512). External amine and guanidine derivatives selectively enhanced the activity of the K512A mutant and the R507A mutant respectively. Analysis of the pH dependence of chemical rescue of the K512A mutant by propargylamine suggested that unprotonated amine in combination with H2PO4-, the protonic form of phosphate presumably utilized in enzymatic ...Continue Reading

References

Apr 1, 1992·Protein Science : a Publication of the Protein Society·M A PhillipsW J Rutter
Jul 7, 1999·The Biochemical Journal·C Eis, B Nidetzky
Jun 29, 2000·Annual Review of Biochemistry·H D Ly, S G Withers
Sep 26, 2000·The Journal of Biological Chemistry·P L AbdianR A Geremia
Sep 28, 2000·The Journal of Biological Chemistry·D M WilliamsP A Cole
Sep 10, 2002·Journal of Molecular Biology·S GeremiaL N Johnson
Apr 15, 2003·Journal of Molecular Biology·Pedro M CoutinhoBernard Henrissat
Mar 23, 2004·Biochemical and Biophysical Research Communications·Alejandra YepJack Preiss
Oct 19, 2004·Chemical Communications : Chem Comm·Luke L Lairson, Stephen G Withers
Jun 14, 2005·Nature Structural & Molecular Biology·James FlintGideon J Davies
Aug 6, 2005·Journal of Molecular Biology·Laurent LarivièreSolange Moréra
Dec 14, 2005·Biochemistry·Yollete V Guillén Schlippe, Lizbeth Hedstrom
Mar 15, 1999·Angewandte Chemie·Tom D Heightman, Andrea T Vasella

❮ Previous
Next ❯

Citations

Apr 9, 2008·The Journal of Biological Chemistry·Matthew W VettingJohn S Blanchard
Mar 11, 2011·Natural Product Reports·Christiane Luley-Goedl, Bernd Nidetzky

❮ 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.