, Volume 68, Issue 4, pp 1103–1113 | Cite as

Endoplasmic reticulium protein profiling of heat-stressed Jurkat cells by one dimensional electrophoresis and liquid chromatography tandem mass spectrometry

  • Xiulian Zhang
  • Yasuhiro Kuramitsu
  • Aiguo Ma
  • Hui Zhang
  • Kazuyuki Nakamura
Original Research


Proteomic study on membrane-integrated proteins in endoplasmic reticulum (ER) fractions was performed. In this study, we examined the effects of heat stress on Jurkat cells. The ER fractions were highly purified by differential centrifugation with sodium carbonate washing and acetone methanol precipitations. The ER membrane proteins were separated by one dimensional electrophoresis (1-DE), and some of the protein bands changed their abundance by heat stress, 12 of the 14 bands containing 40 and 60 ribosomal proteins whose expression level were decreased, on the contrary, 2 of the 14 bands containing ubiquitin and eukaryotic translation initiation factor 3 were increased. Heat treatment of human Jurkat cells led to an increase in the phosphorylation of PERK and eIF2α within 30 min of exposure. This was followed by an increase in the expression of the GRP78. Protein ubiquitination and subsequent degradation by the proteasome are important mechanisms regulating cell cycle, growth and differentiation, the result showed that heat stress enhanced ubiquitination modification of the microsomal proteins. The data of this study strongly suggest that heat treatment led to a significant reduction in protein expression and activated UPR, concomitant with protein hyperubiqutination in ER.


Endoplasmic reticulium Heat stress SDS-PAGE LC–MS/MS 



Endoplasmic reticulium


60S ribosomal protein L19

CBB R-250

Coomassie brilliant blue R-250


Sodium lauryl sulfate


Polyacrylamide gel electrophoresis




Tris buffered saline



MPI preparation was supported by Prof. Bill Jordan (Center for Biodiscovery and School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand).

Conflict of interest

The authors declared that they have no competing interests.


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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Xiulian Zhang
    • 1
    • 2
  • Yasuhiro Kuramitsu
    • 3
  • Aiguo Ma
    • 1
  • Hui Zhang
    • 4
  • Kazuyuki Nakamura
    • 3
  1. 1.The Institute of Human NutritionMedical College of Qingdao UniversityQingdaoPeople’s Republic of China
  2. 2.Qingdao Center for Disease Control and PreventionQingdaoPeople’s Republic of China
  3. 3.Department of Biochemistry and Functional ProteomicsYamaguchi University Graduate School of MedicineUbeJapan
  4. 4.Zaozhuang University HospitalZaozhuangPeople’s Republic of China

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