Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form: Molecular basis of enzyme deficiency

Ferenc Orosz, Judit Oláh, Marco Alvarez, György M. Keserü, Beáta Szabó, Gábor Wágner, Zoltán Kovári, Margit Horányi, Klára Baróti, Joseph A. Martial, Susan Hollán, Judit Ovádi

Resultado de la investigación: Article

26 Citas (Scopus)

Resumen

In a Hungarian family with severe decrease in triosephosphate isomerase (TPI) activity, 2 germ line-identical but phenotypically differing compound heterozygote brothers inherited 2 independent (Phe240Leu and Glu145stop codon) mutations. The kinetic, thermodynamic, and associative properties of the recombinant human wild-type and Phe240Leu mutant enzymes were compared with those of TPIs in normal and deficient erythrocyte hemolysates. The specific activity of the recombinant mutant enzyme relative to the wild type was much higher (30%) than expected from the activity (3%) measured in hemolysates. Enhanced attachment of mutant TPI to erythrocyte inside-out vesicles and to microtubules of brain cells was found when the binding was measured with TPIs in hemolysate. In contrast, there was no difference between the binding of the recombinant wild-type and Phe240Leu mutant enzymes. These findings suggest that the missense mutation by itself is not enough to explain the low catalytic activity and "stickiness" of mutant TPI observed in hemolysate. The activity of the mutant TPI is further reduced by its attachment to inside-out vesicles or microtubules. Comparative studies of the hemolysate from a British patient with Glu104Asp homozygosity and with the platelet lysates from the Hungarian family suggest that the microcompartmentation of TPI is not unique for the hemolysates from the Hungarian TPI-deficient brothers. The possible role of cellular components, other than the mutant enzymes, in the distinct behavior of TPI in isolated form versus in hemolysates from the compound heterozygotes and the simple heterozygote family members is discussed.

Idioma originalEnglish
Páginas (desde-hasta)3106-3112
Número de páginas7
PublicaciónBlood
Volumen98
N.º10
DOI
EstadoPublished - 15 nov 2001

Huella dactilar

Triose-Phosphate Isomerase
Enzymes
Heterozygote
Microtubules
Siblings
Erythrocytes
Missense Mutation
Platelets
Thermodynamics
Germ Cells
Codon
Catalyst activity
Brain
Blood Platelets
Mutation
Kinetics

ASJC Scopus subject areas

  • Immunology
  • Biochemistry
  • Hematology
  • Cell Biology

Citar esto

Orosz, Ferenc ; Oláh, Judit ; Alvarez, Marco ; Keserü, György M. ; Szabó, Beáta ; Wágner, Gábor ; Kovári, Zoltán ; Horányi, Margit ; Baróti, Klára ; Martial, Joseph A. ; Hollán, Susan ; Ovádi, Judit. / Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form : Molecular basis of enzyme deficiency. En: Blood. 2001 ; Vol. 98, N.º 10. pp. 3106-3112.
@article{553909fac43f44fc80ce7f1a87bc4c73,
title = "Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form: Molecular basis of enzyme deficiency",
abstract = "In a Hungarian family with severe decrease in triosephosphate isomerase (TPI) activity, 2 germ line-identical but phenotypically differing compound heterozygote brothers inherited 2 independent (Phe240Leu and Glu145stop codon) mutations. The kinetic, thermodynamic, and associative properties of the recombinant human wild-type and Phe240Leu mutant enzymes were compared with those of TPIs in normal and deficient erythrocyte hemolysates. The specific activity of the recombinant mutant enzyme relative to the wild type was much higher (30{\%}) than expected from the activity (3{\%}) measured in hemolysates. Enhanced attachment of mutant TPI to erythrocyte inside-out vesicles and to microtubules of brain cells was found when the binding was measured with TPIs in hemolysate. In contrast, there was no difference between the binding of the recombinant wild-type and Phe240Leu mutant enzymes. These findings suggest that the missense mutation by itself is not enough to explain the low catalytic activity and {"}stickiness{"} of mutant TPI observed in hemolysate. The activity of the mutant TPI is further reduced by its attachment to inside-out vesicles or microtubules. Comparative studies of the hemolysate from a British patient with Glu104Asp homozygosity and with the platelet lysates from the Hungarian family suggest that the microcompartmentation of TPI is not unique for the hemolysates from the Hungarian TPI-deficient brothers. The possible role of cellular components, other than the mutant enzymes, in the distinct behavior of TPI in isolated form versus in hemolysates from the compound heterozygotes and the simple heterozygote family members is discussed.",
author = "Ferenc Orosz and Judit Ol{\'a}h and Marco Alvarez and Keser{\"u}, {Gy{\"o}rgy M.} and Be{\'a}ta Szab{\'o} and G{\'a}bor W{\'a}gner and Zolt{\'a}n Kov{\'a}ri and Margit Hor{\'a}nyi and Kl{\'a}ra Bar{\'o}ti and Martial, {Joseph A.} and Susan Holl{\'a}n and Judit Ov{\'a}di",
year = "2001",
month = "11",
day = "15",
doi = "10.1182/blood.V98.10.3106",
language = "English",
volume = "98",
pages = "3106--3112",
journal = "Blood",
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Orosz, F, Oláh, J, Alvarez, M, Keserü, GM, Szabó, B, Wágner, G, Kovári, Z, Horányi, M, Baróti, K, Martial, JA, Hollán, S & Ovádi, J 2001, 'Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form: Molecular basis of enzyme deficiency', Blood, vol. 98, n.º 10, pp. 3106-3112. https://doi.org/10.1182/blood.V98.10.3106

Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form : Molecular basis of enzyme deficiency. / Orosz, Ferenc; Oláh, Judit; Alvarez, Marco; Keserü, György M.; Szabó, Beáta; Wágner, Gábor; Kovári, Zoltán; Horányi, Margit; Baróti, Klára; Martial, Joseph A.; Hollán, Susan; Ovádi, Judit.

En: Blood, Vol. 98, N.º 10, 15.11.2001, p. 3106-3112.

Resultado de la investigación: Article

TY - JOUR

T1 - Distinct behavior of mutant triosephosphate isomerase in hemolysate and in isolated form

T2 - Molecular basis of enzyme deficiency

AU - Orosz, Ferenc

AU - Oláh, Judit

AU - Alvarez, Marco

AU - Keserü, György M.

AU - Szabó, Beáta

AU - Wágner, Gábor

AU - Kovári, Zoltán

AU - Horányi, Margit

AU - Baróti, Klára

AU - Martial, Joseph A.

AU - Hollán, Susan

AU - Ovádi, Judit

PY - 2001/11/15

Y1 - 2001/11/15

N2 - In a Hungarian family with severe decrease in triosephosphate isomerase (TPI) activity, 2 germ line-identical but phenotypically differing compound heterozygote brothers inherited 2 independent (Phe240Leu and Glu145stop codon) mutations. The kinetic, thermodynamic, and associative properties of the recombinant human wild-type and Phe240Leu mutant enzymes were compared with those of TPIs in normal and deficient erythrocyte hemolysates. The specific activity of the recombinant mutant enzyme relative to the wild type was much higher (30%) than expected from the activity (3%) measured in hemolysates. Enhanced attachment of mutant TPI to erythrocyte inside-out vesicles and to microtubules of brain cells was found when the binding was measured with TPIs in hemolysate. In contrast, there was no difference between the binding of the recombinant wild-type and Phe240Leu mutant enzymes. These findings suggest that the missense mutation by itself is not enough to explain the low catalytic activity and "stickiness" of mutant TPI observed in hemolysate. The activity of the mutant TPI is further reduced by its attachment to inside-out vesicles or microtubules. Comparative studies of the hemolysate from a British patient with Glu104Asp homozygosity and with the platelet lysates from the Hungarian family suggest that the microcompartmentation of TPI is not unique for the hemolysates from the Hungarian TPI-deficient brothers. The possible role of cellular components, other than the mutant enzymes, in the distinct behavior of TPI in isolated form versus in hemolysates from the compound heterozygotes and the simple heterozygote family members is discussed.

AB - In a Hungarian family with severe decrease in triosephosphate isomerase (TPI) activity, 2 germ line-identical but phenotypically differing compound heterozygote brothers inherited 2 independent (Phe240Leu and Glu145stop codon) mutations. The kinetic, thermodynamic, and associative properties of the recombinant human wild-type and Phe240Leu mutant enzymes were compared with those of TPIs in normal and deficient erythrocyte hemolysates. The specific activity of the recombinant mutant enzyme relative to the wild type was much higher (30%) than expected from the activity (3%) measured in hemolysates. Enhanced attachment of mutant TPI to erythrocyte inside-out vesicles and to microtubules of brain cells was found when the binding was measured with TPIs in hemolysate. In contrast, there was no difference between the binding of the recombinant wild-type and Phe240Leu mutant enzymes. These findings suggest that the missense mutation by itself is not enough to explain the low catalytic activity and "stickiness" of mutant TPI observed in hemolysate. The activity of the mutant TPI is further reduced by its attachment to inside-out vesicles or microtubules. Comparative studies of the hemolysate from a British patient with Glu104Asp homozygosity and with the platelet lysates from the Hungarian family suggest that the microcompartmentation of TPI is not unique for the hemolysates from the Hungarian TPI-deficient brothers. The possible role of cellular components, other than the mutant enzymes, in the distinct behavior of TPI in isolated form versus in hemolysates from the compound heterozygotes and the simple heterozygote family members is discussed.

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U2 - 10.1182/blood.V98.10.3106

DO - 10.1182/blood.V98.10.3106

M3 - Article

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AN - SCOPUS:0035892130

VL - 98

SP - 3106

EP - 3112

JO - Blood

JF - Blood

SN - 0006-4971

IS - 10

ER -