Endoplasmic Reticulum-Mediated Protein Quality Control and Endoplasmic Reticulum-Associated Degradation Pathway Explain the Reduction of N-glycoprotein Level Under the Lead Stress

Du, Hong; Zheng, Canqi; Aslam, Muhmmad; Xie, Xihui; Wang, Wanna; Yang, Yingquan; Liu, Xiaojuan

doi:10.3389/fpls.2020.598552

Cited by 5 publications

(8 citation statements)

References 75 publications

(99 reference statements)

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“…If their normal transport can be partially restored by overexpression of related chaperones, the mutant protein can still perform a compensatory function ( 26 ). In other words, the upstream regulatory mechanism of HERG channel abnormalities is the ERQC system, which mediates channel protein folding and transport in the ER, and serves a vital role in changes in ion channel function ( 27 ). The aforementioned viewpoints were verified in the present study.…”

Section: Discussionmentioning

confidence: 99%

Role and mechanism of chaperones calreticulin and ERP57 in restoring trafficking to mutant HERG‑A561V protein

Huang

Zheng

et al. 2021

Int J Mol Med

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Long QT syndrome type 2 is caused by a mutation in the human-ether-a-go-go-related gene (HERG) gene encoding the rapidly activating delayed rectifier K-current. HERG is a key cell membrane glycoprotein; however, whether the maturation process of HERG protein involves key molecules derived from the calnexin (cNX)/calreticulin (cRT) cycle and how these molecules work remains unknown. Using western blotting, the present study screened the key molecules CNX/CRT/endoplasmic reticulum protein 57 (ERP57) involved in this cycle, and it was revealed that the protein expression levels of cNX/cRT/ERP57 in wild-type (WT)/A561V cells were increased compared with those in WT cells (n=3; P<0.05). Additionally, a co-immunoprecipitation experiment was used to reveal that the ability of cNX/ERP57/cRT to interact with HERG was significantly increased in A561V and WT/A561V cells (n=3; P<0.05). A plasmid lacking the bb' domain of ERP57 was constructed and it was demonstrated that the key site of ERP57 binding to CRT and immature HERG protein is the bb' domain. The whole-cell patch-clamp technique detected that the tail current density increased by 46% following overexpression of cRT and by 53% following overexpression of ERP57 in WT/A561V cells. Overexpression of CRT and ERP57 could increased HERG protein levels on the membrane detected by confocal imaging. Furthermore, overexpression of ERP57 and cRT proteins could restore the HERG-A561V mutant protein trafficking process and rescue the dominant-negative suppression of WT. Overall, ERP57/CRT served a crucial role in the HERG-A561V mutant protein trafficking deficiency and degradation process.

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Section: Discussionmentioning

confidence: 99%

Role and mechanism of chaperones calreticulin and ERP57 in restoring trafficking to mutant HERG‑A561V protein

Huang

Zheng

et al. 2021

Int J Mol Med

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“…The precipitation was washed three times using the cold acetone and then dissolved in 8 M urea. The protein concentration was measured with BCA kit according to the instructions of manufacturer (ab102536, Abcam, China) ( Du et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

Global Profiling of N-Glycoproteins and N-Glycans in the Diatom Phaeodactylum tricornutum

Xie

Chen

et al. 2021

Front. Plant Sci.

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N-glycosylation is an important posttranslational modification in all eukaryotes, but little is known about the N-glycoproteins and N-glycans in microalgae. Here, N-glycoproteomic and N-glycomic approaches were used to unveil the N-glycoproteins and N-glycans in the model diatom Phaeodactylum tricornutum. In total, 863 different N-glycopeptides corresponding to 639 N-glycoproteins were identified from P. tricornutum. These N-glycoproteins participated in a variety of important metabolic pathways in P. tricornutum. Twelve proteins participating in the N-glycosylation pathway were identified as N-glycoproteins, indicating that the N-glycosylation of these proteins might be important for the protein N-glycosylation pathway. Subsequently, 69 N-glycans corresponding to 59 N-glycoproteins were identified and classified into high mannose and hybrid type N-glycans. High mannose type N-glycans contained four different classes, such as Man-5, Man-7, Man-9, and Man-10 with a terminal glucose residue. Hybrid type N-glycan harbored Man-4 with a terminal GlcNAc residue. The identification of N-glycosylation on nascent proteins expanded our understanding of this modification at a N-glycoproteomic scale, the analysis of N-glycan structures updated the N-glycan database in microalgae. The results obtained from this study facilitate the elucidation of the precise function of these N-glycoproteins and are beneficial for future designing the microalga to produce the functional humanized biopharmaceutical N-glycoproteins for the clinical therapeutics.

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“…Cell density, Fv/Fm, the activity of peroxide dismutase (POD) and superoxide dismutase (SOD), the content of malondialdehyde (MDA) in P. tricornutum were measured as described in our previous paper [31]. The soluble protein was analyzed via Coomassie brilliant blue kit (Jiancheng Biotech Company, Nanjing, China) via UV-VIS spectrophotometry at 595 nm [6]. The soluble sugar was analyzed by the methods of [13] using UV-VIS spectrophotometry at 490 nm.…”

Section: Measurements Of Physiological Parametersmentioning

confidence: 99%

“…Based on previous papers and bioinformatic analysis in silico, the working model of ERQC pathway was proposed (Fig. 5) [1,6,14]. After the inhibition of tunicamycin on the first step of protein N-glycosylation modification, the expression of ALG genes, STT3 subunit of OST, CRT and PDI genes were upregulated to orchestrate the protein N-glycosylation modification.…”

Section: Gene Id Abbreviationmentioning

confidence: 99%

“…It was known that GnTI was responsible for the formation of complex-type N-glycan structure in Due to the differentially expressed genes during the pathway of protein N-glycosylation, it is interesting to know the N-glycan structure under tunicamycin stress. So far, the N-glycan structures were widely identified in animals and higher plants, however, the relative information in microalgae is very limit [6]. The N-glycan structures in diatom P. tricornutum were previously reported to be mannose-type ranging from Man-5 to Man-9 via MALDI-TOF MS, IMS-MS and ESI-MS n fragmentation patterns data [1,7,17].…”

Section: Gene Id Abbreviationmentioning

confidence: 99%

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Endoplasmic reticulum-quality control pathway and endoplasmic reticulum-associated degradation mechanism regulate the N-glycoproteins and N-glycan structures in the diatom Phaeodactylum tricornutum

Chen

Liu

et al. 2022

Microb Cell Fact

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Tunicamycin inhibits the first step of protein N-glycosylation modification. However, the physiological, transcriptomic, and N-glycomic effects of tunicamycin on important marine diatom Phaeodactylum tricornutum are still unknown. In this study, comprehensive approaches were used to study the effects of tunicamycin stress. The results showed that cell growth and photosynthesis were significantly inhibited in P. tricornutum under the tunicamycin stress. The soluble protein content was significantly decreased, while the soluble sugar and neutral lipid were dramatically increased to orchestrate the balance of carbon and nitrogen metabolisms. The stress of 0.3 μg ml−1 tunicamycin resulted in the differential expression of ERQC and ERAD related genes. The upregulation of genes involved in ERQC pathway, the activation of anti-oxidases and the differential expression of genes related with ERAD mechanism might be important for maintaining homeostasis in cell. The identification of N-glycans, especially complex-type N-glycan structures enriched the N-glycan database of diatom P. tricornutum and provided important information for studying the function of N-glycosylation modification on proteins. As a whole, our study proposed working models of ERQC and ERAD will provide a solid foundation for further in-depth study of the related mechanism and the diatom expression system.

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Endoplasmic Reticulum-Mediated Protein Quality Control and Endoplasmic Reticulum-Associated Degradation Pathway Explain the Reduction of N-glycoprotein Level Under the Lead Stress

Cited by 5 publications

References 75 publications

Role and mechanism of chaperones calreticulin and ERP57 in restoring trafficking to mutant HERG‑A561V protein

Role and mechanism of chaperones calreticulin and ERP57 in restoring trafficking to mutant HERG‑A561V protein

Global Profiling of N-Glycoproteins and N-Glycans in the Diatom Phaeodactylum tricornutum

Endoplasmic reticulum-quality control pathway and endoplasmic reticulum-associated degradation mechanism regulate the N-glycoproteins and N-glycan structures in the diatom Phaeodactylum tricornutum

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