scholarly journals The neutron structure of Leishmania mexicana triose phosphate isomerase with transition state mimics reveals general base catalyst

2019 ◽  
Vol 75 (a2) ◽  
pp. e114-e114
Author(s):  
Vinardas Kelpsas ◽  
Octav Caldararu ◽  
Yashraj Kulkarni ◽  
Rikkert Wierenga ◽  
Ulf Ryde ◽  
...  
Keyword(s):  
Transition State ◽  
Triose Phosphate Isomerase ◽  
Base Catalyst ◽  
Leishmania Mexicana ◽  
Triose Phosphate ◽  
General Base ◽  
Neutron Structure

scholarly journals The ionization of a buried glutamic acid is thermodynamically linked to the stability of Leishmania mexicana triose phosphate isomerase

2000 ◽  
Vol 267 (9) ◽  
pp. 2516-2524 ◽  
Author(s):  
Anne-Marie Lambeir ◽  
Jan Backmann ◽  
Javier Ruiz-Sanz ◽  
Vladimir Filimonov ◽  
Jens Erik Nielsen ◽  
...  
Keyword(s):  
Glutamic Acid ◽  
Triose Phosphate Isomerase ◽  
Leishmania Mexicana ◽  
Triose Phosphate ◽  
The Stability

scholarly journals Perdeuteration, large crystal growth and neutron data collection ofLeishmania mexicanatriose-phosphate isomerase E65Q variant

2019 ◽  
Vol 75 (4) ◽  
pp. 260-269 ◽  
Author(s):  
Vinardas Kelpšas ◽  
Bénédicte Lafumat ◽  
Matthew P. Blakeley ◽  
Nicolas Coquelle ◽  
Esko Oksanen ◽  
...  
Keyword(s):  
Triose Phosphate Isomerase ◽  
Dihydroxyacetone Phosphate ◽  
Leishmania Mexicana ◽  
Large Crystal ◽  
Neutron Data ◽  
X Ray ◽  
Triose Phosphate ◽  
Catalytic Mechanisms ◽  
Crystal Volume ◽  
Neutron Crystallography

Triose-phosphate isomerase (TIM) catalyses the interconversion of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. Two catalytic mechanisms have been proposed based on two reaction-intermediate analogues, 2-phosphoglycolate (2PG) and phosphoglycolohydroxamate (PGH), that have been used as mimics of thecis-enediol(ate) intermediate in several studies of TIM. The protonation states that are critical for the mechanistic interpretation of these structures are generally not visible in the X-ray structures. To resolve these questions, it is necessary to determine the hydrogen positions using neutron crystallography. Neutron crystallography requires large crystals and benefits from replacing all hydrogens with deuterium.Leishmania mexicanatriose-phosphate isomerase was therefore perdeuterated and large crystals with 2PG and PGH were produced. Neutron diffraction data collected from two crystals with different volumes highlighted the importance of crystal volume, as smaller crystals required longer exposures and resulted in overall worse statistics.

scholarly journals Evidence for lysine 80 as general base catalyst of leucine dehydrogenase.

1993 ◽  
Vol 268 (36) ◽  
pp. 27039-27045
Author(s):  
T Sekimoto ◽  
T Matsuyama ◽  
T Fukui ◽  
K Tanizawa
Keyword(s):  
Base Catalyst ◽  
Leucine Dehydrogenase ◽  
General Base

scholarly journals A High-Content Screening Assay for Small Molecules That Stabilize Mutant Triose Phosphate Isomerase (TPI) as Treatments for TPI Deficiency

2021 ◽  
pp. 247255522110181
Author(s):  
Andreas Vogt ◽  
Samantha L. Eicher ◽  
Tracey D. Myers ◽  
Stacy L. Hrizo ◽  
Laura L. Vollmer ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Large Scale ◽  
Protein Quality ◽  
Fluorescent Protein ◽  
Triose Phosphate Isomerase ◽  
Pharmacological Intervention ◽  
Screening Assay ◽  
Triose Phosphate ◽  
The Common ◽  
Green Fluorescent

Triose phosphate isomerase deficiency (TPI Df) is an untreatable, childhood-onset glycolytic enzymopathy. Patients typically present with frequent infections, anemia, and muscle weakness that quickly progresses with severe neuromusclar dysfunction requiring aided mobility and often respiratory support. Life expectancy after diagnosis is typically ~5 years. There are several described pathogenic mutations that encode functional proteins; however, these proteins, which include the protein resulting from the “common” TPIE105D mutation, are unstable due to active degradation by protein quality control (PQC) pathways. Previous work has shown that elevating mutant TPI levels by genetic or pharmacological intervention can ameliorate symptoms of TPI Df in fruit flies. To identify compounds that increase levels of mutant TPI, we have developed a human embryonic kidney (HEK) stable knock-in model expressing the common TPI Df protein fused with green fluorescent protein (HEK TPIE105D-GFP). To directly address the need for lead TPI Df therapeutics, these cells were developed into an optical drug discovery platform that was implemented for high-throughput screening (HTS) and validated in 3-day variability tests, meeting HTS standards. We initially used this assay to screen the 446-member National Institutes of Health (NIH) Clinical Collection and validated two of the hits in dose–response, by limited structure–activity relationship studies with a small number of analogs, and in an orthogonal, non-optical assay in patient fibroblasts. The data form the basis for a large-scale phenotypic screening effort to discover compounds that stabilize TPI as treatments for this devastating childhood disease.

scholarly journals Triose Phosphate Isomerase from Bakers' Yeast

1972 ◽  
Vol 247 (10) ◽  
pp. 3361-3362
Author(s):  
Stuart W. Hawkinson ◽  
Chin Hsuan Wei ◽  
Fred C. Hartman ◽  
I. Lucille Norton ◽  
J. Ralph Einstein
Keyword(s):  
Triose Phosphate Isomerase ◽  
Triose Phosphate
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