Incorporation of ortho- and meta-tyrosine into cellular proteins leads to erythropoietin-resistance in an erythroid cell line

Esztella Mikolás, Szilárd Kun, Boglárka Laczy, Gergo A. Molnár, Eszter Sélley, T. Kőszegi, I. Wittmann

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background/Aims: Erythropoietin-resistance is an unsolved concern in the treatment of renal anaemia. We aimed to investigate the possible role of ortho- and meta-tyrosine - the hydroxyl free radical products of L-phenylalanine - in the development of erythropoietin-resistance. Methods: TF-1 erythroblast cell line was used. Cell concentration was determined on day 1; 2 and 3 by two independent observers simultaneously in Bürker cell counting chambers. Protein concentration was determined with colorimetric method. Para-, ortho- and meta-tyrosine levels were measured using reverse phase-HPLC with fluorescence detection. Using Western blot method activating phosphorylation of STAT5 and ERK1/2 were investigated. Results: We found a time- and concentration-dependent decrease of erythropoietin-induced proliferative activity in case of ortho- and meta-tyrosine treated TF-1 erythroblasts, compared to the para-tyrosine cultured cells. Decreased erythropoietin-response could be regained with a competitive dose of para-tyrosine. Proteins of erythroblasts treated by ortho- or meta-tyrosine had lower para-tyrosine and higher ortho- or meta-tyrosine content. Activating phosphorylation of ERK and STAT5 due to erythropoietin was practically prevented by ortho- or meta-tyrosine treatment. Conclusion: According to this study elevated ortho- and meta-tyrosine content of erythroblasts may lead to the dysfunction of intracellular signaling, resulting in erythropoietin-hyporesponsiveness.

Original languageEnglish
Pages (from-to)217-225
Number of pages9
JournalKidney and Blood Pressure Research
Volume38
Issue number2-3
DOIs
Publication statusPublished - 2014

Fingerprint

Erythroid Cells
Erythropoietin
Erythroblasts
Cell Line
Proteins
Tyrosine
Phosphorylation
3-tyrosine
2-tyrosine
Phenylalanine
Hydroxyl Radical
Free Radicals
Anemia
Cultured Cells
Fluorescence
Western Blotting
High Pressure Liquid Chromatography
Kidney

Keywords

  • Cell signaling
  • Erythropoietin-resistance
  • Oxidative stress
  • Tyrosine isomers

ASJC Scopus subject areas

  • Nephrology
  • Cardiology and Cardiovascular Medicine
  • Medicine(all)

Cite this

Incorporation of ortho- and meta-tyrosine into cellular proteins leads to erythropoietin-resistance in an erythroid cell line. / Mikolás, Esztella; Kun, Szilárd; Laczy, Boglárka; Molnár, Gergo A.; Sélley, Eszter; Kőszegi, T.; Wittmann, I.

In: Kidney and Blood Pressure Research, Vol. 38, No. 2-3, 2014, p. 217-225.

Research output: Contribution to journalArticle

Mikolás, Esztella ; Kun, Szilárd ; Laczy, Boglárka ; Molnár, Gergo A. ; Sélley, Eszter ; Kőszegi, T. ; Wittmann, I. / Incorporation of ortho- and meta-tyrosine into cellular proteins leads to erythropoietin-resistance in an erythroid cell line. In: Kidney and Blood Pressure Research. 2014 ; Vol. 38, No. 2-3. pp. 217-225.
@article{6a3e2ce6c4b347fa95e49c09b046dec9,
title = "Incorporation of ortho- and meta-tyrosine into cellular proteins leads to erythropoietin-resistance in an erythroid cell line",
abstract = "Background/Aims: Erythropoietin-resistance is an unsolved concern in the treatment of renal anaemia. We aimed to investigate the possible role of ortho- and meta-tyrosine - the hydroxyl free radical products of L-phenylalanine - in the development of erythropoietin-resistance. Methods: TF-1 erythroblast cell line was used. Cell concentration was determined on day 1; 2 and 3 by two independent observers simultaneously in B{\"u}rker cell counting chambers. Protein concentration was determined with colorimetric method. Para-, ortho- and meta-tyrosine levels were measured using reverse phase-HPLC with fluorescence detection. Using Western blot method activating phosphorylation of STAT5 and ERK1/2 were investigated. Results: We found a time- and concentration-dependent decrease of erythropoietin-induced proliferative activity in case of ortho- and meta-tyrosine treated TF-1 erythroblasts, compared to the para-tyrosine cultured cells. Decreased erythropoietin-response could be regained with a competitive dose of para-tyrosine. Proteins of erythroblasts treated by ortho- or meta-tyrosine had lower para-tyrosine and higher ortho- or meta-tyrosine content. Activating phosphorylation of ERK and STAT5 due to erythropoietin was practically prevented by ortho- or meta-tyrosine treatment. Conclusion: According to this study elevated ortho- and meta-tyrosine content of erythroblasts may lead to the dysfunction of intracellular signaling, resulting in erythropoietin-hyporesponsiveness.",
keywords = "Cell signaling, Erythropoietin-resistance, Oxidative stress, Tyrosine isomers",
author = "Esztella Mikol{\'a}s and Szil{\'a}rd Kun and Bogl{\'a}rka Laczy and Moln{\'a}r, {Gergo A.} and Eszter S{\'e}lley and T. Kőszegi and I. Wittmann",
year = "2014",
doi = "10.1159/000355770",
language = "English",
volume = "38",
pages = "217--225",
journal = "Kidney and Blood Pressure Research",
issn = "1420-4096",
publisher = "S. Karger AG",
number = "2-3",

}

TY - JOUR

T1 - Incorporation of ortho- and meta-tyrosine into cellular proteins leads to erythropoietin-resistance in an erythroid cell line

AU - Mikolás, Esztella

AU - Kun, Szilárd

AU - Laczy, Boglárka

AU - Molnár, Gergo A.

AU - Sélley, Eszter

AU - Kőszegi, T.

AU - Wittmann, I.

PY - 2014

Y1 - 2014

N2 - Background/Aims: Erythropoietin-resistance is an unsolved concern in the treatment of renal anaemia. We aimed to investigate the possible role of ortho- and meta-tyrosine - the hydroxyl free radical products of L-phenylalanine - in the development of erythropoietin-resistance. Methods: TF-1 erythroblast cell line was used. Cell concentration was determined on day 1; 2 and 3 by two independent observers simultaneously in Bürker cell counting chambers. Protein concentration was determined with colorimetric method. Para-, ortho- and meta-tyrosine levels were measured using reverse phase-HPLC with fluorescence detection. Using Western blot method activating phosphorylation of STAT5 and ERK1/2 were investigated. Results: We found a time- and concentration-dependent decrease of erythropoietin-induced proliferative activity in case of ortho- and meta-tyrosine treated TF-1 erythroblasts, compared to the para-tyrosine cultured cells. Decreased erythropoietin-response could be regained with a competitive dose of para-tyrosine. Proteins of erythroblasts treated by ortho- or meta-tyrosine had lower para-tyrosine and higher ortho- or meta-tyrosine content. Activating phosphorylation of ERK and STAT5 due to erythropoietin was practically prevented by ortho- or meta-tyrosine treatment. Conclusion: According to this study elevated ortho- and meta-tyrosine content of erythroblasts may lead to the dysfunction of intracellular signaling, resulting in erythropoietin-hyporesponsiveness.

AB - Background/Aims: Erythropoietin-resistance is an unsolved concern in the treatment of renal anaemia. We aimed to investigate the possible role of ortho- and meta-tyrosine - the hydroxyl free radical products of L-phenylalanine - in the development of erythropoietin-resistance. Methods: TF-1 erythroblast cell line was used. Cell concentration was determined on day 1; 2 and 3 by two independent observers simultaneously in Bürker cell counting chambers. Protein concentration was determined with colorimetric method. Para-, ortho- and meta-tyrosine levels were measured using reverse phase-HPLC with fluorescence detection. Using Western blot method activating phosphorylation of STAT5 and ERK1/2 were investigated. Results: We found a time- and concentration-dependent decrease of erythropoietin-induced proliferative activity in case of ortho- and meta-tyrosine treated TF-1 erythroblasts, compared to the para-tyrosine cultured cells. Decreased erythropoietin-response could be regained with a competitive dose of para-tyrosine. Proteins of erythroblasts treated by ortho- or meta-tyrosine had lower para-tyrosine and higher ortho- or meta-tyrosine content. Activating phosphorylation of ERK and STAT5 due to erythropoietin was practically prevented by ortho- or meta-tyrosine treatment. Conclusion: According to this study elevated ortho- and meta-tyrosine content of erythroblasts may lead to the dysfunction of intracellular signaling, resulting in erythropoietin-hyporesponsiveness.

KW - Cell signaling

KW - Erythropoietin-resistance

KW - Oxidative stress

KW - Tyrosine isomers

UR - http://www.scopus.com/inward/record.url?scp=84898674100&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84898674100&partnerID=8YFLogxK

U2 - 10.1159/000355770

DO - 10.1159/000355770

M3 - Article

C2 - 24751667

AN - SCOPUS:84898674100

VL - 38

SP - 217

EP - 225

JO - Kidney and Blood Pressure Research

JF - Kidney and Blood Pressure Research

SN - 1420-4096

IS - 2-3

ER -