Characterization of distinct heat shock- and thapsigargin-induced cytoprotective proteins in FRTL-5 thyroid cells

Robert Christian Smallridge, Irene D. Gist, George C. Tsokos, Juliann G. Kiang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Heat shock induces the expression of proteins with molecular weights of 70-72 kd and 90 kd, whereas thapsigargin induces the expression of a glucose- regulated protein 78 kd (GRP-78) in certain cells. In this study we examined the induction and cytoprotective effects of heat shock- and thapsigargin- induced proteins in FRTL-5 rat thyroid cells. New protein synthesis was assessed in [35S]methionine-labeled cells and quantitated densitometrically. The expression of specific stress proteins was identified using Western blots, whereas cytoprotection provided by these proteins was evaluated by trypan blue exclusion. Exposure to heat shock (45°C, 15 minutes) induced the expression of proteins with molecular weights at the range of low 70 kD and low 90 kD that peaked between 2-6 hours and returned to baseline within 24 hours. Treatment of cells with thapsigargin (200 nM, 15 minutes) induced the expression of different molecular weight proteins, most likely GRP-78 and -94, that peaked at 4-6 hours and lasted for 24 hours. Neither the removal of growth factors (thyroid-stimulating hormone and insulin) for 5 days nor the elimination of extracellular Ca2+ with EGTA or clamping of the intracellular Ca2+ with BAPTA for 15 minutes affected expression of the heat shock- and the thapsigargin-induced stress proteins. In contrast, protein kinase C inhibitors H7 and GF109203X abolished the expression of all three groups of stress proteins. Both heat shock- and thapsigargin-induced proteins completely protected cells from subsequent thermal injury (47°C, 35 minutes). The induction of cytoprotective proteins by heat shock and thapsigargin requires the presence of protein kinase C but is Ca2+- and growth factor-independent.

Original languageEnglish (US)
Pages (from-to)1041-1047
Number of pages7
JournalThyroid
Volume9
Issue number10
StatePublished - Oct 1999

Fingerprint

Thapsigargin
Shock
Thyroid Gland
Hot Temperature
Heat-Shock Proteins
Proteins
Molecular Weight
Protein Kinase C
Intercellular Signaling Peptides and Proteins
Cytoprotection
Trypan Blue
Protein C Inhibitor
Egtazic Acid
Thyrotropin
Protein Kinase Inhibitors
Constriction
Methionine
Western Blotting
Insulin
Wounds and Injuries

ASJC Scopus subject areas

  • Endocrinology

Cite this

Characterization of distinct heat shock- and thapsigargin-induced cytoprotective proteins in FRTL-5 thyroid cells. / Smallridge, Robert Christian; Gist, Irene D.; Tsokos, George C.; Kiang, Juliann G.

In: Thyroid, Vol. 9, No. 10, 10.1999, p. 1041-1047.

Research output: Contribution to journalArticle

Smallridge, Robert Christian ; Gist, Irene D. ; Tsokos, George C. ; Kiang, Juliann G. / Characterization of distinct heat shock- and thapsigargin-induced cytoprotective proteins in FRTL-5 thyroid cells. In: Thyroid. 1999 ; Vol. 9, No. 10. pp. 1041-1047.
@article{5caeecdde4274fcc972e84b2b87869d6,
title = "Characterization of distinct heat shock- and thapsigargin-induced cytoprotective proteins in FRTL-5 thyroid cells",
abstract = "Heat shock induces the expression of proteins with molecular weights of 70-72 kd and 90 kd, whereas thapsigargin induces the expression of a glucose- regulated protein 78 kd (GRP-78) in certain cells. In this study we examined the induction and cytoprotective effects of heat shock- and thapsigargin- induced proteins in FRTL-5 rat thyroid cells. New protein synthesis was assessed in [35S]methionine-labeled cells and quantitated densitometrically. The expression of specific stress proteins was identified using Western blots, whereas cytoprotection provided by these proteins was evaluated by trypan blue exclusion. Exposure to heat shock (45°C, 15 minutes) induced the expression of proteins with molecular weights at the range of low 70 kD and low 90 kD that peaked between 2-6 hours and returned to baseline within 24 hours. Treatment of cells with thapsigargin (200 nM, 15 minutes) induced the expression of different molecular weight proteins, most likely GRP-78 and -94, that peaked at 4-6 hours and lasted for 24 hours. Neither the removal of growth factors (thyroid-stimulating hormone and insulin) for 5 days nor the elimination of extracellular Ca2+ with EGTA or clamping of the intracellular Ca2+ with BAPTA for 15 minutes affected expression of the heat shock- and the thapsigargin-induced stress proteins. In contrast, protein kinase C inhibitors H7 and GF109203X abolished the expression of all three groups of stress proteins. Both heat shock- and thapsigargin-induced proteins completely protected cells from subsequent thermal injury (47°C, 35 minutes). The induction of cytoprotective proteins by heat shock and thapsigargin requires the presence of protein kinase C but is Ca2+- and growth factor-independent.",
author = "Smallridge, {Robert Christian} and Gist, {Irene D.} and Tsokos, {George C.} and Kiang, {Juliann G.}",
year = "1999",
month = "10",
language = "English (US)",
volume = "9",
pages = "1041--1047",
journal = "Thyroid",
issn = "1050-7256",
publisher = "Mary Ann Liebert Inc.",
number = "10",

}

TY - JOUR

T1 - Characterization of distinct heat shock- and thapsigargin-induced cytoprotective proteins in FRTL-5 thyroid cells

AU - Smallridge, Robert Christian

AU - Gist, Irene D.

AU - Tsokos, George C.

AU - Kiang, Juliann G.

PY - 1999/10

Y1 - 1999/10

N2 - Heat shock induces the expression of proteins with molecular weights of 70-72 kd and 90 kd, whereas thapsigargin induces the expression of a glucose- regulated protein 78 kd (GRP-78) in certain cells. In this study we examined the induction and cytoprotective effects of heat shock- and thapsigargin- induced proteins in FRTL-5 rat thyroid cells. New protein synthesis was assessed in [35S]methionine-labeled cells and quantitated densitometrically. The expression of specific stress proteins was identified using Western blots, whereas cytoprotection provided by these proteins was evaluated by trypan blue exclusion. Exposure to heat shock (45°C, 15 minutes) induced the expression of proteins with molecular weights at the range of low 70 kD and low 90 kD that peaked between 2-6 hours and returned to baseline within 24 hours. Treatment of cells with thapsigargin (200 nM, 15 minutes) induced the expression of different molecular weight proteins, most likely GRP-78 and -94, that peaked at 4-6 hours and lasted for 24 hours. Neither the removal of growth factors (thyroid-stimulating hormone and insulin) for 5 days nor the elimination of extracellular Ca2+ with EGTA or clamping of the intracellular Ca2+ with BAPTA for 15 minutes affected expression of the heat shock- and the thapsigargin-induced stress proteins. In contrast, protein kinase C inhibitors H7 and GF109203X abolished the expression of all three groups of stress proteins. Both heat shock- and thapsigargin-induced proteins completely protected cells from subsequent thermal injury (47°C, 35 minutes). The induction of cytoprotective proteins by heat shock and thapsigargin requires the presence of protein kinase C but is Ca2+- and growth factor-independent.

AB - Heat shock induces the expression of proteins with molecular weights of 70-72 kd and 90 kd, whereas thapsigargin induces the expression of a glucose- regulated protein 78 kd (GRP-78) in certain cells. In this study we examined the induction and cytoprotective effects of heat shock- and thapsigargin- induced proteins in FRTL-5 rat thyroid cells. New protein synthesis was assessed in [35S]methionine-labeled cells and quantitated densitometrically. The expression of specific stress proteins was identified using Western blots, whereas cytoprotection provided by these proteins was evaluated by trypan blue exclusion. Exposure to heat shock (45°C, 15 minutes) induced the expression of proteins with molecular weights at the range of low 70 kD and low 90 kD that peaked between 2-6 hours and returned to baseline within 24 hours. Treatment of cells with thapsigargin (200 nM, 15 minutes) induced the expression of different molecular weight proteins, most likely GRP-78 and -94, that peaked at 4-6 hours and lasted for 24 hours. Neither the removal of growth factors (thyroid-stimulating hormone and insulin) for 5 days nor the elimination of extracellular Ca2+ with EGTA or clamping of the intracellular Ca2+ with BAPTA for 15 minutes affected expression of the heat shock- and the thapsigargin-induced stress proteins. In contrast, protein kinase C inhibitors H7 and GF109203X abolished the expression of all three groups of stress proteins. Both heat shock- and thapsigargin-induced proteins completely protected cells from subsequent thermal injury (47°C, 35 minutes). The induction of cytoprotective proteins by heat shock and thapsigargin requires the presence of protein kinase C but is Ca2+- and growth factor-independent.

UR - http://www.scopus.com/inward/record.url?scp=0032712685&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032712685&partnerID=8YFLogxK

M3 - Article

C2 - 10560962

AN - SCOPUS:0032712685

VL - 9

SP - 1041

EP - 1047

JO - Thyroid

JF - Thyroid

SN - 1050-7256

IS - 10

ER -