Crystal structures of two monomeric triosephosphate isomerase variants identifiedviaa directed-evolution protocol selecting forL-arabinose isomerase activity
نویسندگان
چکیده
منابع مشابه
Structure-based directed evolution of a monomeric triosephosphate isomerase: toward a pentose sugar isomerase.
Through structure-based and directed evolution approaches, a new catalytic activity has been established on the (β/α)8 barrel enzyme triosephosphate isomerase (TIM). This work started from ml8bTIM, a monomeric variant of TIM, in which the phosphate-binding loop (loop-8) had been shortened. Structure analysis suggested an additional point mutation (V233A), converting ml8bTIM into A-TIM. A-TIM ha...
متن کاملDesign, creation, and characterization of a stable, monomeric triosephosphate isomerase.
Protein engineering on trypanosomal triosephosphate isomerase (TIM) converted this oligomeric enzyme into a stable, monomeric protein that is enzymatically active. Wild-type TIM consists of two identical subunits that form a very tight dimer involving interactions of 32 residues of each subunit. By replacing 15 residues of the major interface loop by another 8-residue fragment, a variant was co...
متن کاملCrystal structure of triosephosphate isomerase from Trypanosoma cruzi in hexane.
To gain insight into the mechanisms of enzyme catalysis in organic solvents, the x-ray structure of some monomeric enzymes in organic solvents was determined. However, it remained to be explored whether the structure of oligomeric proteins is also amenable to such analysis. The field acquired new perspectives when it was proposed that the x-ray structure of enzymes in nonaqueous media could rev...
متن کاملThe mechanism of the triosephosphate isomerase reaction.
The interconversion of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate catalyzed by the enzyme triosephosphate isomerase was first demonstrated by Meyerhof and Kiessling (1). Since then studies with the enzyme have disclosed little about the mechanism of this reaction. Such an interconversion may be envisioned as occurring by formation of an enediol as, based on chemical analogy (2)) ...
متن کاملTriosephosphate isomerase by consensus design: dramatic differences in physical properties and activity of related variants.
Consensus design, the selection of mutations based on the most common amino acid in each position of a multiple sequence alignment, has proven to be an efficient way to engineer stabilized mutants and even to design entire proteins. However, its application has been limited to small motifs or small families of highly related proteins. Also, we have little idea of how information that specifies ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Acta Crystallographica Section F Structural Biology Communications
سال: 2016
ISSN: 2053-230X
DOI: 10.1107/s2053230x16007548