FEBS LettersPub Date : 2021-03-01Epub Date: 2020-10-07DOI: 10.1002/1873-3468.13937
Ludwik Gorczyca, Jianyao Du, Kristin M Bircsak, Xia Wen, Anna M Vetrano, Lauren M Aleksunes
{"title":"Low oxygen tension differentially regulates the expression of placental solute carriers and ABC transporters.","authors":"Ludwik Gorczyca, Jianyao Du, Kristin M Bircsak, Xia Wen, Anna M Vetrano, Lauren M Aleksunes","doi":"10.1002/1873-3468.13937","DOIUrl":"https://doi.org/10.1002/1873-3468.13937","url":null,"abstract":"Low oxygen concentration, or hypoxia, is an important physiological regulator of placental function including chemical disposition. Here, we compared the ability of low oxygen tension to alter the expression of solute carriers (SLC) and ABC transporters in two human placental models, namely BeWo cells and term placental explants. We found that exposure to low oxygen concentration differentially regulates transporter expression in BeWo cells, including downregulation of ENT1, OATP4A1, OCTN2, BCRP, and MRP2/3/5, and upregulation of CNT1, OAT4, OATP2B1, SERT, SOAT, and MRP1. Similar upregulation of MRP1 and downregulation of MRP5 and BCRP were observed in explants, whereas uptake transporters were decreased or unchanged. Furthermore, a screening of transcriptional regulators of transporters revealed that hypoxia leads to a decrease in the mRNA levels of aryl hydrocarbon receptor, nuclear factor erythroid 2‐related factor 2, and retinoid x receptor alpha in both human placental models. These data suggest that transporter expression is differentially regulated by oxygen concentration across experimental human placental models.","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 6","pages":"811-827"},"PeriodicalIF":3.5,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13937","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38522451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2021-03-01Epub Date: 2020-11-05DOI: 10.1002/1873-3468.13957
Flora Szeri, Fatemeh Niaziorimi, Sylvia Donnelly, Joseph Orndorff, Koen van de Wetering
{"title":"Generation of fully functional fluorescent fusion proteins to gain insights into ABCC6 biology.","authors":"Flora Szeri, Fatemeh Niaziorimi, Sylvia Donnelly, Joseph Orndorff, Koen van de Wetering","doi":"10.1002/1873-3468.13957","DOIUrl":"10.1002/1873-3468.13957","url":null,"abstract":"<p><p>ABCC6 mediates release of ATP from hepatocytes into the blood. Extracellularly, ATP is converted into the mineralization inhibitor pyrophosphate. Consequently, inactivating mutations in ABCC6 give low plasma pyrophosphate and underlie the ectopic mineralization disorder pseudoxanthoma elasticum. How ABCC6 mediates cellular ATP release is still unknown. Fluorescent ABCC6 fusion proteins would allow mechanistic studies, but fluorophores attached to the ABCC6 N- or C-terminus result in intracellular retention and degradation. Here we describe that intramolecular introduction of fluorophores yields fully functional ABCC6 fusion proteins. A corresponding ABCC6 variant in which the catalytic glutamate of the second nucleotide binding domain was mutated, correctly routed to the plasma membrane but was inactive. Finally, N-terminal His<sup>10</sup> or FLAG tags did not affect activity of the fusion proteins, allowing their purification for biochemical characterization.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 6","pages":"799-810"},"PeriodicalIF":3.5,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7987643/pdf/nihms-1638648.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38592541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2021-03-01Epub Date: 2020-11-11DOI: 10.1002/1873-3468.13974
Carlo W T van Roermund, Lodewijk IJlst, Alison Baker, Ronald J A Wanders, Freddie L Theodoulou, Hans R Waterham
{"title":"The Saccharomyces cerevisiae ABC subfamily D transporter Pxa1/Pxa2p co-imports CoASH into the peroxisome.","authors":"Carlo W T van Roermund, Lodewijk IJlst, Alison Baker, Ronald J A Wanders, Freddie L Theodoulou, Hans R Waterham","doi":"10.1002/1873-3468.13974","DOIUrl":"https://doi.org/10.1002/1873-3468.13974","url":null,"abstract":"<p><p>ATP-binding cassette (ABC) subfamily D transporters are important for the uptake of fatty acids and other beta-oxidation substrates into peroxisomes. Genetic and biochemical evidence indicates that the transporters accept fatty acyl-coenzyme A that is cleaved during the transport cycle and then re-esterified in the peroxisomal lumen. However, it is not known whether free coenzyme A (CoA) is released inside or outside the peroxisome. Here we have used Saccharomyces cerevisiae and isolated peroxisomes to demonstrate that free CoA is released in the peroxisomal lumen. Thus, ABC subfamily D transporter provide an import pathway for free CoA that controls peroxisomal CoA homeostasis and tunes metabolism according to the cell's demands.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 6","pages":"763-772"},"PeriodicalIF":3.5,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13974","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38541542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2021-03-01Epub Date: 2020-10-20DOI: 10.1002/1873-3468.13950
Maki Tsujita, Boris Vaisman, Liu Chengyu, Kasey C Vickers, Kei-Ichiro Okuhira, Sten Braesch-Andersen, Alan T Remaley
{"title":"Apolipoprotein A-I in mouse cerebrospinal fluid derives from the liver and intestine via plasma high-density lipoproteins assembled by ABCA1 and LCAT.","authors":"Maki Tsujita, Boris Vaisman, Liu Chengyu, Kasey C Vickers, Kei-Ichiro Okuhira, Sten Braesch-Andersen, Alan T Remaley","doi":"10.1002/1873-3468.13950","DOIUrl":"https://doi.org/10.1002/1873-3468.13950","url":null,"abstract":"<p><p>Apolipoprotein (apo) A-I, the major structural protein of high-density lipoprotein (HDL), is present in human and mouse cerebrospinal fluid (CSF) despite its lack of expression in brain cells. To identify the origin of apoA-I in CSF, we generated intestine-specific and liver-specific Apoa1 knockout mice (Apoa1<sup>ΔInt</sup> and Apoa1<sup>Δliv</sup> mice, respectively). Lipoprotein profiles of Apoa1<sup>ΔInt</sup> and Apoa1<sup>ΔLiv</sup> mice resembled those of control littermates, whereas knockout of Apoa1 in both intestine and liver (Apoa1<sup>ΔIntΔLiv</sup> ) resulted in a 60-percent decrease in HDL-cholesterol levels, thus strongly mimicking the Apoa1<sup>-/-</sup> mice. Immunoassays revealed that mouse apoA-I was not present in the CSF of the Apoa1<sup>ΔIntΔLiv</sup> mice. Furthermore, apoA-I levels in CSF were highly correlated with plasma spherical HDL levels, which were regulated by ABCA1 and LCAT. Collectively, these results suggest that apoA-I protein in CSF originates in liver and small intestine and is taken up from the plasma.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 6","pages":"773-788"},"PeriodicalIF":3.5,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13950","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38457774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2021-03-01Epub Date: 2020-11-28DOI: 10.1002/1873-3468.13991
Eszter Kozák, Bence Szikora, Attila Iliás, Péter K Jani, Zoltán Hegyi, Zsolt Matula, Dóra Dedinszki, Natália Tőkési, Krisztina Fülöp, Viola Pomozi, György Várady, Éva Bakos, Gabor E Tusnády, Imre Kacskovics, Andras Váradi
{"title":"Creation of the first monoclonal antibody recognizing an extracellular epitope of hABCC6.","authors":"Eszter Kozák, Bence Szikora, Attila Iliás, Péter K Jani, Zoltán Hegyi, Zsolt Matula, Dóra Dedinszki, Natália Tőkési, Krisztina Fülöp, Viola Pomozi, György Várady, Éva Bakos, Gabor E Tusnády, Imre Kacskovics, Andras Váradi","doi":"10.1002/1873-3468.13991","DOIUrl":"https://doi.org/10.1002/1873-3468.13991","url":null,"abstract":"<p><p>Mutations in the ABCC6 gene result in calcification diseases such as pseudoxanthoma elasticum or Generalized Arterial Calcification of Infancy. Generation of antibodies recognizing an extracellular (EC) epitope of ABCC6 has been hampered by the short EC segments of the protein. To overcome this limitation, we immunized bovine FcRn transgenic mice exhibiting an augmented humoral immune response with Human Embryonic Kidney 293 cells cells expressing human ABCC6 (hABCC6). We obtained a monoclonal antibody recognizing an EC epitope of hABCC6 that we named mEChC6. Limited proteolysis revealed that the epitope is within a loop in the N-terminal half of ABCC6 and probably spans amino acids 338-347. mEChC6 recognizes hABCC6 in the liver of hABCC6 transgenic mice, verifying both specificity and EC binding to intact hepatocytes.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 6","pages":"789-798"},"PeriodicalIF":3.5,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13991","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38577380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Dynamics and electrostatics define an allosteric druggable site within the receptor-binding domain of SARS-CoV-2 spike protein.","authors":"Sayan Bhattacharjee, Rajanya Bhattacharyya, Jayati Sengupta","doi":"10.1002/1873-3468.14038","DOIUrl":"https://doi.org/10.1002/1873-3468.14038","url":null,"abstract":"<p><p>The pathogenesis of the SARS-CoV-2 virus initiates through recognition of the angiotensin-converting enzyme 2 (ACE2) receptor of the host cells by the receptor-binding domain (RBD) located at the spikes of the virus. Here, using molecular dynamics simulations, we have demonstrated the allosteric crosstalk within the RBD in the apo- and the ACE2 receptor-bound states, revealing the contribution of the dynamics-based correlated motions and the electrostatic energy perturbations to this crosstalk. While allostery, based on correlated motions, dominates inherent distal communication in the apo-RBD, the electrostatic energy perturbations determine favorable pairwise crosstalk within the RBD residues upon binding to ACE2. Interestingly, the allosteric path is composed of residues which are evolutionarily conserved within closely related coronaviruses, pointing toward the biological relevance of the communication and its potential as a target for drug development.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 4","pages":"442-451"},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38824053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Halophilic to mesophilic adaptation of ubiquitin-like proteins.","authors":"Quan Li, Mengqing Li, Cong Li, Xinxin Li, Chenghui Lu, Xiaoming Tu, Zhiyong Zhang, Xuecheng Zhang","doi":"10.1002/1873-3468.14023","DOIUrl":"https://doi.org/10.1002/1873-3468.14023","url":null,"abstract":"Elucidating how proteins adapt from halophilic to mesophilic environments will enable a better understanding of protein evolution and folding. In this study, by directed evolution and site‐directed mutagenesis of the halophilic ubiquitin‐like protein (ULP) Samp2, we find that substitution of the prebiotic amino acid Asp31 by Gly is uniquely effective in the mesophilic adaptation of ULP. Sequence analysis shows that substitution of Asp/Glu in halophilic ULPs by Gly in mesophilic ULPs has higher occurrence than other substitutions, supporting the unique role of the substitution in the mesophilic adaptation of ULP. Molecular dynamic simulations indicate that the mesophilic adaptation might result from the effect of the substitution on the conformational flexibility of ULP.","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 4","pages":"521-531"},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14023","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38697458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Insertion loop-mediated folding propagation governs efficient maturation of hyperthermophilic Tk-subtilisin at high temperatures.","authors":"Ryo Uehara, Nanako Dan, Hiroshi Amesaka, Takuya Yoshizawa, Yuichi Koga, Shigenori Kanaya, Kazufumi Takano, Hiroyoshi Matsumura, Shun-Ichi Tanaka","doi":"10.1002/1873-3468.14028","DOIUrl":"https://doi.org/10.1002/1873-3468.14028","url":null,"abstract":"<p><p>The serine protease Tk-subtilisin from the hyperthermophilic archaeon Thermococcus kodakarensis possesses three insertion loops (IS1-IS3) on its surface, as compared to its mesophilic counterparts. Although IS1 and IS2 are required for maturation of Tk-subtilisin at high temperatures, the role of IS3 remains unknown. Here, CD spectroscopy revealed that IS3 deletion arrested Tk-subtilisin folding at an intermediate state, in which the central nucleus was formed, but the subsequent folding propagation into terminal subdomains did not occur. Alanine substitution of the aspartate residue in IS3 disturbed the intraloop hydrogen-bonding network, as evidenced by crystallographic analysis, resulting in compromised folding at high temperatures. Taking into account the high conservation of IS3 across hyperthermophilic homologues, we propose that the presence of IS3 is important for folding of hyperthermophilic subtilisins in high-temperature environments.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 4","pages":"452-461"},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14028","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38368838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
FEBS LettersPub Date : 2021-02-01Epub Date: 2021-01-24DOI: 10.1002/1873-3468.14029
Jiss Maria Louis, Arjun Agarwal, Raviprasad Aduri, Indrani Talukdar
{"title":"Global analysis of RNA-protein interactions in TNF-α induced alternative splicing in metabolic disorders.","authors":"Jiss Maria Louis, Arjun Agarwal, Raviprasad Aduri, Indrani Talukdar","doi":"10.1002/1873-3468.14029","DOIUrl":"https://doi.org/10.1002/1873-3468.14029","url":null,"abstract":"<p><p>In this report, using the database of RNA-binding protein specificities (RBPDB) and our previously published RNA-seq data, we analyzed the interactions between RNA and RNA-binding proteins to decipher the role of alternative splicing in metabolic disorders induced by TNF-α. We identified 13 395 unique RNA-RBP interactions, including 385 unique RNA motifs and 35 RBPs, some of which (including MBNL-1 and 3, ZFP36, ZRANB2, and SNRPA) are transcriptionally regulated by TNF-α. In addition to some previously reported RBPs, such as RBMX and HuR/ELAVL1, we found a few novel RBPs, such as ZRANB2 and SNRPA, to be involved in the regulation of metabolic syndrome-associated genes that contain an enrichment of tetrameric RNA sequences (AUUU). Taken together, this study paves the way for novel RNA-protein interaction-based therapeutics for treating metabolic syndromes.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":"595 4","pages":"476-490"},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38796930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}