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GRP78 protects CHO cells from ribosylation
Wu, Beibei1; Yu, Lexiang1,5; Hu, Pingdong1,5; Lu, Yang1; Li, Juan2; Wei, Yan1; He, Rongqiao1,2,3,4; Y. Wei; R. He
2018-04-01
Source PublicationBIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
Correspondent Emailyanwei@ibp.ac.cn ; rongqiaohe@163.com
ISSN0167-4889
SubtypeArticle
Volume1865Issue:4Pages:629-637
Contribution Rank1,2
Abstract

D-Ribose (Rib), a reactive glycation compound that exists in organisms, abnormally increases in the urine of diabetic patients and can yield large amounts of advanced glycation end products (AGEs), leading to cell dysfunction. However, whether cellular proteins are sensitive to this type of glycation is unknown. In this study, we found that cellular AGEs accumulate in Chinese hamster ovary (CHO) cells with increased Rib concentration and administration time. Mass spectrum analysis of isolated AGE-modified proteins from cell lysates showed that glucose-regulated protein 78 kD (GRP78) is one of the main ribosylated proteins. Co-immunoprecipitation assays further confirmed the interaction between AGEs and GRP78. Compared with D-glucose (Glc), Rib produced much more AGEs in cells. In kinetic studies, the first order rate constant of LDH released from CHO cells incubated with Rib was nearly 8-fold higher than that of Glc, suggesting that Rib is highly cytotoxic. Immunofluorescent co-localization analysis manifested partial superimposition of AGEs and GRP78, which were distributed throughout the endoplasmic reticulum. Western blotting showed that the expression of GRP78 is up-regulated and then down-regulated in CHO cells during Rib treatment. In the presence of Rib, the suppression of GRP78 expression either with transfected siRNA or with the inhibitor (-)-epigallocatechin gallate (EGCG) dramatically increased AGE levels and decreased cell viability compared with these parameters in the control groups. GRP78 over-expression decreased AGE levels and rescued the cells from Rib-induced cytotoxicity. These data indicate that GRP78 plays a role in preventing Rib-induced CHO cell cytotoxicity.

KeywordD-Ribose GRP78 Glycation AGEs Cytotoxicity
DOI10.1016/j.bbamcr.2018.02.001
Indexed BySCI
Language英语
Funding OrganizationNatural Scientific Foundation of China NSFC(31270868 ; National Key Research and Development Program of China(2016YFC1305900) ; Beijing Municipal Science and Technology Project(Z161100000217141 ; QCAS Biotechnology Fund(GJHZ1131) ; Youth Innovation Promotion Association CAS(2017132) ; 31670805) ; Z161100000216137)
WOS Research AreaBiochemistry & Molecular Biology ; Cell Biology
WOS SubjectBiochemistry & Molecular Biology ; Cell Biology
WOS IDWOS:000427335700009
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS KeywordGLYCATION END-PRODUCTS ; ENDOPLASMIC-RETICULUM STRESS ; GLUCOSE-REGULATED PROTEIN ; ER STRESS ; D-RIBOSE ; CEREBROSPINAL-FLUID ; ACTIVATION ; APOPTOSIS ; INDUCTION ; ACCUMULATION
Citation statistics
Document Type期刊论文
Identifierhttp://ir.psych.ac.cn/handle/311026/26049
Collection脑与认知科学国家重点实验室
Corresponding AuthorY. Wei; R. He
Affiliation1.Chinese Acad Sci, Inst Biophys, State Key Lab Brain & Cognit Sci, 15 Datun Rd, Beijing 100101, Peoples R China
2.Chinese Acad Sci, Inst Psychol, Key Lab Mental Hlth, Beijing 100101, Peoples R China
3.Southwest Med Univ, Luzhou 646000, Sichuan, Peoples R China
4.Capital Med Univ, Beijing Inst Brain Disorders, Alzheimers Dis Ctr, Beijing 10069, Peoples R China
5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Wu, Beibei,Yu, Lexiang,Hu, Pingdong,et al. GRP78 protects CHO cells from ribosylation[J]. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH,2018,1865(4):629-637.
APA Wu, Beibei.,Yu, Lexiang.,Hu, Pingdong.,Lu, Yang.,Li, Juan.,...&R. He.(2018).GRP78 protects CHO cells from ribosylation.BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH,1865(4),629-637.
MLA Wu, Beibei,et al."GRP78 protects CHO cells from ribosylation".BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH 1865.4(2018):629-637.
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