A stress-inducible 40 kDa protein (hsp40): Purification by modified two-dimensional gel electrophoresis and co-localization with hsc70(p73) in heat-shocked HeLa cells

H. Hattori, T. Kaneda, Balakrishna L Lokeshwar, A. Laszlo, K. Ohtsuka

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

We have previously reported that a novel 40 kDa protein is induced by heat shock and several environmental stresses in mammalian and avian cells and that the N-terminal amino acid sequence of this 40 kDa protein has homology with the bacterial DnaJ heat-shock protein. We have purified this protein (40 kDa heat-shock protein, hsp40) from HeLa cells by modified two-dimensional gel electrophoresis and generated a polyclonal antibody against hsp40. This antibody was highly specific for human hsp40 and cross-reacted weakly with rat and Chinese hamster hsp40. Indirect immunofluorescence revealed that the hsp40 in HeLa cells accumulates in the nucleus, especially in the nucleolus, during heat shock and returns to the cytoplasm during the recovery period. The kinetics of the accumulation in the nucleoli and subsequent return to the cytoplasm of hsp40 was similar to that of hsp70. In addition, hsp40 was co-localized with hsc70(p73) in heat-shocked HeLa cells as demonstrated by double immunofluorescence staining. These results suggest that hsp40 (a DnaJ homologue) and hsp70 (a DnaK homologue) may act in concert to repair (refold) denatured proteins and protein aggregates in the nuclei and nucleoli of heat-shocked HeLa cells.

Original languageEnglish (US)
Pages (from-to)629-638
Number of pages10
JournalJournal of Cell Science
Volume104
Issue number3
StatePublished - Jan 1 1993
Externally publishedYes

Fingerprint

Electrophoresis, Gel, Two-Dimensional
HeLa Cells
HSP40 Heat-Shock Proteins
Hot Temperature
Shock
Cytoplasm
Proteins
Protein Refolding
Antibodies
Indirect Fluorescent Antibody Technique
Heat-Shock Proteins
Cricetulus
Fluorescent Antibody Technique
Amino Acid Sequence
Staining and Labeling

Keywords

  • Co-localization
  • hsp40 (DnaJ homologue)
  • hsp70 (DnaK homologue)

ASJC Scopus subject areas

  • Cell Biology

Cite this

A stress-inducible 40 kDa protein (hsp40) : Purification by modified two-dimensional gel electrophoresis and co-localization with hsc70(p73) in heat-shocked HeLa cells. / Hattori, H.; Kaneda, T.; Lokeshwar, Balakrishna L; Laszlo, A.; Ohtsuka, K.

In: Journal of Cell Science, Vol. 104, No. 3, 01.01.1993, p. 629-638.

Research output: Contribution to journalArticle

@article{59aaae5ea5b74a579207c49d65c867ba,
title = "A stress-inducible 40 kDa protein (hsp40): Purification by modified two-dimensional gel electrophoresis and co-localization with hsc70(p73) in heat-shocked HeLa cells",
abstract = "We have previously reported that a novel 40 kDa protein is induced by heat shock and several environmental stresses in mammalian and avian cells and that the N-terminal amino acid sequence of this 40 kDa protein has homology with the bacterial DnaJ heat-shock protein. We have purified this protein (40 kDa heat-shock protein, hsp40) from HeLa cells by modified two-dimensional gel electrophoresis and generated a polyclonal antibody against hsp40. This antibody was highly specific for human hsp40 and cross-reacted weakly with rat and Chinese hamster hsp40. Indirect immunofluorescence revealed that the hsp40 in HeLa cells accumulates in the nucleus, especially in the nucleolus, during heat shock and returns to the cytoplasm during the recovery period. The kinetics of the accumulation in the nucleoli and subsequent return to the cytoplasm of hsp40 was similar to that of hsp70. In addition, hsp40 was co-localized with hsc70(p73) in heat-shocked HeLa cells as demonstrated by double immunofluorescence staining. These results suggest that hsp40 (a DnaJ homologue) and hsp70 (a DnaK homologue) may act in concert to repair (refold) denatured proteins and protein aggregates in the nuclei and nucleoli of heat-shocked HeLa cells.",
keywords = "Co-localization, hsp40 (DnaJ homologue), hsp70 (DnaK homologue)",
author = "H. Hattori and T. Kaneda and Lokeshwar, {Balakrishna L} and A. Laszlo and K. Ohtsuka",
year = "1993",
month = "1",
day = "1",
language = "English (US)",
volume = "104",
pages = "629--638",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "3",

}

TY - JOUR

T1 - A stress-inducible 40 kDa protein (hsp40)

T2 - Purification by modified two-dimensional gel electrophoresis and co-localization with hsc70(p73) in heat-shocked HeLa cells

AU - Hattori, H.

AU - Kaneda, T.

AU - Lokeshwar, Balakrishna L

AU - Laszlo, A.

AU - Ohtsuka, K.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - We have previously reported that a novel 40 kDa protein is induced by heat shock and several environmental stresses in mammalian and avian cells and that the N-terminal amino acid sequence of this 40 kDa protein has homology with the bacterial DnaJ heat-shock protein. We have purified this protein (40 kDa heat-shock protein, hsp40) from HeLa cells by modified two-dimensional gel electrophoresis and generated a polyclonal antibody against hsp40. This antibody was highly specific for human hsp40 and cross-reacted weakly with rat and Chinese hamster hsp40. Indirect immunofluorescence revealed that the hsp40 in HeLa cells accumulates in the nucleus, especially in the nucleolus, during heat shock and returns to the cytoplasm during the recovery period. The kinetics of the accumulation in the nucleoli and subsequent return to the cytoplasm of hsp40 was similar to that of hsp70. In addition, hsp40 was co-localized with hsc70(p73) in heat-shocked HeLa cells as demonstrated by double immunofluorescence staining. These results suggest that hsp40 (a DnaJ homologue) and hsp70 (a DnaK homologue) may act in concert to repair (refold) denatured proteins and protein aggregates in the nuclei and nucleoli of heat-shocked HeLa cells.

AB - We have previously reported that a novel 40 kDa protein is induced by heat shock and several environmental stresses in mammalian and avian cells and that the N-terminal amino acid sequence of this 40 kDa protein has homology with the bacterial DnaJ heat-shock protein. We have purified this protein (40 kDa heat-shock protein, hsp40) from HeLa cells by modified two-dimensional gel electrophoresis and generated a polyclonal antibody against hsp40. This antibody was highly specific for human hsp40 and cross-reacted weakly with rat and Chinese hamster hsp40. Indirect immunofluorescence revealed that the hsp40 in HeLa cells accumulates in the nucleus, especially in the nucleolus, during heat shock and returns to the cytoplasm during the recovery period. The kinetics of the accumulation in the nucleoli and subsequent return to the cytoplasm of hsp40 was similar to that of hsp70. In addition, hsp40 was co-localized with hsc70(p73) in heat-shocked HeLa cells as demonstrated by double immunofluorescence staining. These results suggest that hsp40 (a DnaJ homologue) and hsp70 (a DnaK homologue) may act in concert to repair (refold) denatured proteins and protein aggregates in the nuclei and nucleoli of heat-shocked HeLa cells.

KW - Co-localization

KW - hsp40 (DnaJ homologue)

KW - hsp70 (DnaK homologue)

UR - http://www.scopus.com/inward/record.url?scp=0027311871&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027311871&partnerID=8YFLogxK

M3 - Article

C2 - 8314866

AN - SCOPUS:0027311871

VL - 104

SP - 629

EP - 638

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 3

ER -