ArticleTraffic (Copenhagen, Denmark)2013
Analysis of ER resident proteins in Saccharomyces cerevisiae: implementation of H/KDEL retrieval sequences.
Article in Traffic (Copenhagen, Denmark), 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 21 citations in OpenAlex.
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- The toxicity of neodymium and genome-scale genetic screen of neodymium-sensitive gene deletion mutations in the yeast Saccharomyces cerevisiae.Environmental science and pollution research international · 2022Article
- Rapid transient expression of functional human vascular endothelial growth factor inBiotechnology reports (Amsterdam, Netherlands) · 2020Article
- Stepwise engineering ofRSC advances · 2019Article
- Yeast Expression Systems: Overview and Recent Advances.Molecular biotechnology · 2019Review
- Silica-based solid-phase extraction of cross-linked nucleic acid-bound proteins.Life science alliance · 2018Article
- The antifungal plant defensin AhPDF1.1b is a beneficial factor involved in adaptive response to zinc overload when it is expressed in yeast cells.MicrobiologyOpen · 2015Article
- FAD-linked Presenilin-1 V97L mutation impede tranport regulation and intracellular Ca(2+) homeostasis under ER stress.International journal of clinical and experimental medicine · 2015Article
- Protein folding and secretion: mechanistic insights advancing recombinant protein production in S. cerevisiae.Current opinion in biotechnology · 2014Review
- BiP clustering facilitates protein folding in the endoplasmic reticulum.PLoS computational biology · 2014Article
- Approaches to imaging unfolded secretory protein stress in living cells.Endoplasmic reticulum stress in diseases · 2014Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
An elaborate quality control system regulates endoplasmic reticulum (ER) homeostasis by ensuring the fidelity of protein synthesis and maturation. In budding yeast, genomic analyses and high-throughput proteomic studies have identified ER resident proteins that restore homeostasis following local perturbations. Yet, how these folding factors modulate stress has been largely unexplored. In this study, we designed a series of polymerase chain reaction (PCR)-based modules including codon-optimized epitopes and fluorescent protein (FP) variants complete with C-terminal H/KDEL retrieval motifs. These conserved sequences are inherent to most soluble ER resident proteins. To monitor multiple proteins simultaneously, H/KDEL cassettes are available with six different selection markers, providing optimal flexibility for live-cell imaging and multicolor labeling in vivo. A single pair of PCR primers can be used for the amplification of these 26 modules, enabling numerous combinations of tags and selection markers. The versatility of pCY H/KDEL cassettes was demonstrated by labeling BiP/Kar2p, Pdi1p and Scj1p with all novel tags, thus providing a direct comparison among FP variants. Furthermore, to advance in vitro studies of yeast ER proteins, Strep-tag II was engineered with a C-terminal retrieval sequence. Here, an efficient purification strategy was established for BiP under physiological conditions.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.