Triose phosphate isomerase from the coelacanth. An approach to the rapid determination of an amino acid sequence with small amounts of material

Kolb, E.D.; Harris, J. Ieuan; Bridgen, John

doi:10.1042/bj1370185

Cited by 85 publications

(22 citation statements)

References 31 publications

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“…The functional importance of Glu-104 to TPI structure and activity is implicated by its conservation in the enzymes of all species that have been characterized to date, including Bacillus stearothermophilus (36), Escherichia coli (37), Saccharomyces cerevisiae (38), coelacanth (39), chicken (40,41), rabbit (42), and human (2,3). Approximately 25% of the TPI amino acid sequence is invariant in all of these species.…”

Section: Discussionmentioning

confidence: 99%

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

Daar¹,

Artymiuk²,

Phillips³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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Triose-phosphate isomerase (TPI; D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1) deficiency is a recessive disorder that results in hemolytic anemia and neuromuscular dysfunction. To determine the molecular basis of this disorder, a TPI allele from two unrelated patients homozygous for TPI deficiency was compared with an allele from a normal individual. Each disease-associated sequence harbors a G-C -+ C-G transversion in the codon for amino acid-104 and specifies a structurally altered protein in which a glutamate residue is replaced by an aspartate residue. The importance of glutamate-104 to enzyme structure and function is implicated by its conservation in the TPI protein of all species that have been characterized to date. The glutamate-toaspartate substitution results in a thermolabile enzyme as demonstrated by assaysof-TPI activity in cultured fibroblasts of each patient and cultured Chinese hamster ovary (CHO) cells that were stably transformed with the mutant alleles. Although this substitution conserves the overall charge of amino acid-104, the x-ray crystal structure of chicken TPI indicates that the loss of a side-chain methylene group (-CH2CH2COO -* -CH2COO ) is sufficient to disrupt the counterbalancing of charges that normally exists within a hydrophobic pocket of the native enzyme.Triose-phosphate isomerase (TPI; D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1) catalyzes the movement of a single proton to interconvert dihydroxyacetone phosphate and glyceraldehyde 3-phosphate in glycolysis and gluconeogenesis (1). The enzyme, a dimer of identical subunits that are 248 amino acids in humans (2, 3), has no cofactors, required metal ions, or cooperativity between subunits. Both prokaryotic and eukaryotic TPI are very similar in sequence and structure and are characterized by a high catalytic efficiency. The second-order rate constant for the formation of dihydroxyacetone phosphate, the thermodynamically favored reaction, is close to the rate constant for a diffusion-limited encounter of substrate and enzyme (4, 5).Site-directed mutagenesis combined with crystallographic and kinetic studies have provided a means of defining the contribution of specific TPI amino acids to enzyme activity (6, 7). This method has focused on amino acids implicated by high-resolution structural data and chemical-modification experiments to be important to the catalytic mechanism of the enzyme. We have chosen a complementary approach to understand TPI structure-function relationships in studying natural TPI gene mutations that exist within the human population. Hereditary TPI deficiency is an autosomal recessive disease that has severe clinical manifestations including chronic hemolytic anemia and neuromuscular disorders (8). Homozygous-deficient individuals usually have 3-20% of normal TPI activity; all of this activity is heat labile, suggesting that at least one allele encodes a structurally altered protein (3, 9, 10). The other allele is presumably null, since mutations producing no detectable ...

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Section: Discussionmentioning

confidence: 99%

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

Daar¹,

Artymiuk²,

Phillips³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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“…Sequence alignment by using the program GCG (Genetics Computer Group, Inc., Copyright 1982, Wisconsin, USA) of all known TIM sequences. In order: Copja, Coptis japonica (Okada et al, 1989); Maize, Zea mays (maize) (Marchionni & Gilbert, 1986); Human, Homo sapiens (Maquat et al, 1985); Macmu, Macacu mulatta (rhesus macaque) (Old & Mohrenweiser, 1988); Rabit, Oryctolagus cuniculus (rabbit) ; Mouse, Mus musculus (mouse) (Cheng et al, 1990); Chick, Callus gallus (chicken) ; Latch, Latimeria chalumnae (latimeria) (Kolb et al, 1974); Culta, Culex tarsalis (mosquito) (Tittiger et al, 1993); Drome, Drosophila melanogaster (fruit fly) (Shaw-Lee et al, 1991); Emeni, Emericella niduluns (Aspergillus nidulans) (McKnight et al, 1986); Schpo, Schizosaccharomyces pombe (fission yeast) (Russell, 1985); Yeast, Saccharomyces cerevisiae (baker's yeast) (Alber & Kawasaki, 1982); Trybb, Trypanosoma brucei brucei (Swinkels et al, 1986); Bacst, Bacillusstearothermophilus (Rentier-Delrue et al, 1993); Ecoli, Escherichia co[i (Pichersky et al, 1984); Corgl, Corynebacterium glutamicum (Eikmanns, 1992) 2.8-A structure of B. stearothermophilus TIM are shown in Table l . There are eight residues lying outside the allowed region in the Ramachandran plot (Fig.…”

Section: Quality Of the Structurementioning

confidence: 99%

Crystal structure of recombinant triosephosphate isomerase from bacillus stearothermophilus. An analysis of potential thermostability factors in six isomerases with known three‐dimensional structures points to the importance of hydrophobic interactions

et al. 1995

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The structure of the thermostable triosephosphate isomerase (TIM) from Bacillus stearothermophilus complexed with the competitive inhibitor 2-phosphoglycolate was determined by X-ray crystallography to a resolution of 2.8 A. The structure was solved by molecular replacement using XPLOR. Twofold averaging and solvent flattening was applied to improve the quality of the map. Active sites in both the subunits are occupied by the inhibitor and the flexible loop adopts the "closed" conformation in either subunit. The crystallographic R-factor is 17.6% with good geometry. The two subunits have an RMS deviation of 0.29 A for 248 C" atoms and have average temperature factors of 18.9 and 15.9 A*, respectively. In both subunits, the active site Lys 10 adopts an unusual $,$ combination.A comparison between the six known thermophilic and mesophilic TIM structures was conducted in order to understand the higher stability of B. stearothermophilus TIM. Although the ratio Arg/(Arg+Lys) is higher in B. stearotherrnophilus TIM, the structure comparisons do not directly correlate this higher ratio to the better stability of the B. stearothermophilus enzyme. A higher number of prolines contributes to the higher stability of B. stearothermophilus TIM. Analysis of the known TIM sequences points out that the replacement of a structurally crucial asparagine by a histidine at the interface of monomers, thus avoiding the risk of deamidation and thereby introducing a negative charge at the interface, may be one of the factors for adaptability at higher temperatures in the TIM family. Analysis of buried cavities and the areas lining these cavities also contributes to the greater thermal stability of the B. stearotherrnophilus enzyme. However, the most outstanding result of the structure comparisons appears to point to the hydrophobic stabilization of dimer formation by burying the largest amount of hydrophobic surface area in B. stearothermophilus TIM compared to all five other known TIM structures.

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“…Reaction of glyceraldehyde-3-phosphate dehydrogenase with iodoacetic acid was carried out essentially as described by Kolb et al [6]. Iod0[2-'~C]acetic acid (33 Ci/mol) was obtained from the Radiochemical Centre Ltd (Amersham, Bucks, Great Britain) and was diluted before use with carrier iodoactic acid (recrystallised from hexane) to a final specific radioactivity of I .O Ci/mol.…”

Section: Curboxymetliylutionmentioning

confidence: 99%

D-Glyceraldehyde-3-Phosphate Dehydrogenase. Complete Amino-Acid Sequence of the Enzyme from Bacillus stearothermophilus

Walker

et al. 1980

Eur J Biochem

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1. The complete amino acid sequence of ~-glyceraldehyde-3-phosphate dehydrogenase from the moderate thermophile Bacillus stearotliermopliilus has been determined.2. This has been achieved largely by the automated sequence analysis of large fragments derived by chemical cleavage with cyanogen bromide, BNPS-skatole [the product of reaction between N-bromosuccinimide and 2-( nitrophenyl-sulphenyl)-3-methylindole] and hydroxylamine and enzymic hydrolysis with trypsin at arginine residues.

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Triose phosphate isomerase from the coelacanth. An approach to the rapid determination of an amino acid sequence with small amounts of material

Cited by 85 publications

References 31 publications

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

Human triose-phosphate isomerase deficiency: a single amino acid substitution results in a thermolabile enzyme.

Crystal structure of recombinant triosephosphate isomerase from bacillus stearothermophilus. An analysis of potential thermostability factors in six isomerases with known three‐dimensional structures points to the importance of hydrophobic interactions

D-Glyceraldehyde-3-Phosphate Dehydrogenase. Complete Amino-Acid Sequence of the Enzyme from Bacillus stearothermophilus

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