TPPP/p25: From unfolded protein to misfolding disease: Prediction and experiments

F. Orosz, G. G. Kovács, A. Lehotzky, J. Oláh, O. Vincze, J. Ovádi

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

TPPP/p25, the first representative of a new protein family, identified as a brain-specific unfolded protein induces aberrant microtubule assemblies in vitro, suppresses mitosis in Drosophila embryo and is accumulated in inclusion bodies of human pathological brain tissues. In this paper, we present prediction and additional experimental data that validate TPPP/p25 to be a new member of the "intrinsically unstructured" protein family. The comparison of these characteristics with that of α-synuclein and tau, involved also in neurodegenerative diseases, suggested that although the primary sequences of these proteins are entirely different, there are similarities in their well-defined unstructured segments interrupted by "stabilization centres", phosphorylation and tubulin binding motives. SK-N-MC neuroblastoma cells were transfected with pEGFP-TPPP/p25 construct and a stable clone denoted K4 was selected and used to establish the effect of this unstructured protein on the energy state/metabolism of the cells. Our data by analyzing the mitochondrial membrane polarization by fluorescence microscopy revealed that the high-energy phosphate production in K4 clone is not damaged by the TPPP/p25 expression. Biochemical analysis with cell homogenates provided quantitative data that the ATP level increased 1.5-fold and the activities of hexokinase, glucosephosphate isomerase, phosphofructokinase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were 1.2 to 2.0-fold higher in K4 as compared to the control. Our modelling using these data and rate equations of the individual enzymes suggests that the TPPP/p25 expression stimulates glucose metabolism. At pathological conditions TPPP/p25 is localized in inclusion bodies in multiple system atrophy, it tightly co-localizes with α-synuclein, partially with tubulin and not with vimentin. The previous and the present studies obtained with immunohistochemistry with pathological human brain tissues rendered it possible to classify among pathological inclusions on the basis of immunolabelling of TPPP/p25, and suggest this protein to be a potential linkage between Parkinson's and Alzheimer's diseases.

Original languageEnglish
Pages (from-to)701-711
Number of pages11
JournalBiology of the Cell
Volume96
Issue number9
DOIs
Publication statusPublished - Dec 2004

Fingerprint

Proteostasis Deficiencies
Synucleins
Intrinsically Disordered Proteins
Inclusion Bodies
Tubulin
Brain
Clone Cells
Triose-Phosphate Isomerase
Polarization Microscopy
Glucose-6-Phosphate Isomerase
Multiple System Atrophy
Phosphofructokinases
Protein Unfolding
Glyceraldehyde-3-Phosphate Dehydrogenases
Proteins
Hexokinase
Mitochondrial Membranes
Vimentin
Neuroblastoma
Fluorescence Microscopy

Keywords

  • Energy state
  • Intrinsically unstructured proteins
  • Neurodegeneration

ASJC Scopus subject areas

  • Cell Biology

Cite this

TPPP/p25 : From unfolded protein to misfolding disease: Prediction and experiments. / Orosz, F.; Kovács, G. G.; Lehotzky, A.; Oláh, J.; Vincze, O.; Ovádi, J.

In: Biology of the Cell, Vol. 96, No. 9, 12.2004, p. 701-711.

Research output: Contribution to journalArticle

@article{2471d179addd416ca2aec1a3551c551f,
title = "TPPP/p25: From unfolded protein to misfolding disease: Prediction and experiments",
abstract = "TPPP/p25, the first representative of a new protein family, identified as a brain-specific unfolded protein induces aberrant microtubule assemblies in vitro, suppresses mitosis in Drosophila embryo and is accumulated in inclusion bodies of human pathological brain tissues. In this paper, we present prediction and additional experimental data that validate TPPP/p25 to be a new member of the {"}intrinsically unstructured{"} protein family. The comparison of these characteristics with that of α-synuclein and tau, involved also in neurodegenerative diseases, suggested that although the primary sequences of these proteins are entirely different, there are similarities in their well-defined unstructured segments interrupted by {"}stabilization centres{"}, phosphorylation and tubulin binding motives. SK-N-MC neuroblastoma cells were transfected with pEGFP-TPPP/p25 construct and a stable clone denoted K4 was selected and used to establish the effect of this unstructured protein on the energy state/metabolism of the cells. Our data by analyzing the mitochondrial membrane polarization by fluorescence microscopy revealed that the high-energy phosphate production in K4 clone is not damaged by the TPPP/p25 expression. Biochemical analysis with cell homogenates provided quantitative data that the ATP level increased 1.5-fold and the activities of hexokinase, glucosephosphate isomerase, phosphofructokinase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were 1.2 to 2.0-fold higher in K4 as compared to the control. Our modelling using these data and rate equations of the individual enzymes suggests that the TPPP/p25 expression stimulates glucose metabolism. At pathological conditions TPPP/p25 is localized in inclusion bodies in multiple system atrophy, it tightly co-localizes with α-synuclein, partially with tubulin and not with vimentin. The previous and the present studies obtained with immunohistochemistry with pathological human brain tissues rendered it possible to classify among pathological inclusions on the basis of immunolabelling of TPPP/p25, and suggest this protein to be a potential linkage between Parkinson's and Alzheimer's diseases.",
keywords = "Energy state, Intrinsically unstructured proteins, Neurodegeneration",
author = "F. Orosz and Kov{\'a}cs, {G. G.} and A. Lehotzky and J. Ol{\'a}h and O. Vincze and J. Ov{\'a}di",
year = "2004",
month = "12",
doi = "10.1016/j.biolcel.2004.08.002",
language = "English",
volume = "96",
pages = "701--711",
journal = "Biology of the Cell",
issn = "0248-4900",
publisher = "Portland Press Ltd.",
number = "9",

}

TY - JOUR

T1 - TPPP/p25

T2 - From unfolded protein to misfolding disease: Prediction and experiments

AU - Orosz, F.

AU - Kovács, G. G.

AU - Lehotzky, A.

AU - Oláh, J.

AU - Vincze, O.

AU - Ovádi, J.

PY - 2004/12

Y1 - 2004/12

N2 - TPPP/p25, the first representative of a new protein family, identified as a brain-specific unfolded protein induces aberrant microtubule assemblies in vitro, suppresses mitosis in Drosophila embryo and is accumulated in inclusion bodies of human pathological brain tissues. In this paper, we present prediction and additional experimental data that validate TPPP/p25 to be a new member of the "intrinsically unstructured" protein family. The comparison of these characteristics with that of α-synuclein and tau, involved also in neurodegenerative diseases, suggested that although the primary sequences of these proteins are entirely different, there are similarities in their well-defined unstructured segments interrupted by "stabilization centres", phosphorylation and tubulin binding motives. SK-N-MC neuroblastoma cells were transfected with pEGFP-TPPP/p25 construct and a stable clone denoted K4 was selected and used to establish the effect of this unstructured protein on the energy state/metabolism of the cells. Our data by analyzing the mitochondrial membrane polarization by fluorescence microscopy revealed that the high-energy phosphate production in K4 clone is not damaged by the TPPP/p25 expression. Biochemical analysis with cell homogenates provided quantitative data that the ATP level increased 1.5-fold and the activities of hexokinase, glucosephosphate isomerase, phosphofructokinase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were 1.2 to 2.0-fold higher in K4 as compared to the control. Our modelling using these data and rate equations of the individual enzymes suggests that the TPPP/p25 expression stimulates glucose metabolism. At pathological conditions TPPP/p25 is localized in inclusion bodies in multiple system atrophy, it tightly co-localizes with α-synuclein, partially with tubulin and not with vimentin. The previous and the present studies obtained with immunohistochemistry with pathological human brain tissues rendered it possible to classify among pathological inclusions on the basis of immunolabelling of TPPP/p25, and suggest this protein to be a potential linkage between Parkinson's and Alzheimer's diseases.

AB - TPPP/p25, the first representative of a new protein family, identified as a brain-specific unfolded protein induces aberrant microtubule assemblies in vitro, suppresses mitosis in Drosophila embryo and is accumulated in inclusion bodies of human pathological brain tissues. In this paper, we present prediction and additional experimental data that validate TPPP/p25 to be a new member of the "intrinsically unstructured" protein family. The comparison of these characteristics with that of α-synuclein and tau, involved also in neurodegenerative diseases, suggested that although the primary sequences of these proteins are entirely different, there are similarities in their well-defined unstructured segments interrupted by "stabilization centres", phosphorylation and tubulin binding motives. SK-N-MC neuroblastoma cells were transfected with pEGFP-TPPP/p25 construct and a stable clone denoted K4 was selected and used to establish the effect of this unstructured protein on the energy state/metabolism of the cells. Our data by analyzing the mitochondrial membrane polarization by fluorescence microscopy revealed that the high-energy phosphate production in K4 clone is not damaged by the TPPP/p25 expression. Biochemical analysis with cell homogenates provided quantitative data that the ATP level increased 1.5-fold and the activities of hexokinase, glucosephosphate isomerase, phosphofructokinase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were 1.2 to 2.0-fold higher in K4 as compared to the control. Our modelling using these data and rate equations of the individual enzymes suggests that the TPPP/p25 expression stimulates glucose metabolism. At pathological conditions TPPP/p25 is localized in inclusion bodies in multiple system atrophy, it tightly co-localizes with α-synuclein, partially with tubulin and not with vimentin. The previous and the present studies obtained with immunohistochemistry with pathological human brain tissues rendered it possible to classify among pathological inclusions on the basis of immunolabelling of TPPP/p25, and suggest this protein to be a potential linkage between Parkinson's and Alzheimer's diseases.

KW - Energy state

KW - Intrinsically unstructured proteins

KW - Neurodegeneration

UR - http://www.scopus.com/inward/record.url?scp=10244264868&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10244264868&partnerID=8YFLogxK

U2 - 10.1016/j.biolcel.2004.08.002

DO - 10.1016/j.biolcel.2004.08.002

M3 - Article

C2 - 15567525

AN - SCOPUS:10244264868

VL - 96

SP - 701

EP - 711

JO - Biology of the Cell

JF - Biology of the Cell

SN - 0248-4900

IS - 9

ER -