{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T22:14:10Z","timestamp":1788300850671,"version":"build-2803163510"},"reference-count":49,"publisher":"American Chemical Society (ACS)","issue":"19","license":[{"start":{"date-parts":[[2020,5,6]],"date-time":"2020-05-06T00:00:00Z","timestamp":1588723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/501100004281","name":"Narodowe Centrum Nauki","doi-asserted-by":"publisher","award":["UMO\/2013\/08\/A\/NZ1\/00866"],"award-info":[{"award-number":["UMO\/2013\/08\/A\/NZ1\/00866"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005632","name":"Narodowe Centrum Badan i Rozwoju","doi-asserted-by":"publisher","award":["1\/235773\/19\/NCBR\/2016"],"award-info":[{"award-number":["1\/235773\/19\/NCBR\/2016"]}],"id":[{"id":"10.13039\/501100005632","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,5,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Decapping scavenger enzymes (DcpSs) are important players in mRNA degradation machinery and conserved in eukaryotes. Importantly, human DcpS is the recognized target for spinal muscular atrophy (SMA) and acute myeloid leukemia (AML) therapy, and has recently been connected to development of intellectual disability. Most recombinant DcpSs used in biochemical and biophysical studies are prepared as tagged proteins, with polyhistidine (His-tag) at the N-terminus or C-terminus. Our work is the first report on the parallel characterization of three versions of DcpSs (native and N- or C-terminally tagged) of three species (humans, Caenorhabditis elegans, and Ascaris suum). The native forms of all three enzymes were prepared by N-(His)10 tag cleavage. Protein thermal stability, measured by differential scanning fluorimetry (DSF), was unaffected in the case of native and tagged versions of human and A. suum DcpS; however, the melting temperature (Tm) of C. elagans DcpS of was significantly influenced by the presence of the additional N- or C-tag. To investigate the impact of the tag positioning on the catalytic properties of DcpS, we tested the hydrolytic activity of native DcpS and their His-tagged counterparts toward cap dinucleotides (m7GpppG and m32,2,7GpppG) and m7GDP. The kinetic data indicate that dinucleotide substrates are hydrolyzed with comparable efficiency by native human and A. suum DcpS and their His-tagged forms. In contrast, both His-tagged C. elegans DcpSs exhibited higher activity toward m7GpppG than the native enzyme. m7GDP is resistant to enzymatic cleavage by all three forms of human and nematode DcpS.<\/jats:p>","DOI":"10.1021\/acsomega.0c00304","type":"journal-article","created":{"date-parts":[[2020,5,6]],"date-time":"2020-05-06T20:09:38Z","timestamp":1588795778000},"page":"10759-10766","source":"Crossref","is-referenced-by-count":10,"title":["Effect of the His-Tag Location on Decapping Scavenger\nEnzymes and Their Hydrolytic Activity toward Cap Analogs"],"prefix":"10.1021","volume":"5","author":[{"given":"Aleksandra","family":"Ferenc-Mrozek","sequence":"first","affiliation":[{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]},{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elzbieta","family":"Bojarska","sequence":"additional","affiliation":[{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janusz","family":"Stepinski","sequence":"additional","affiliation":[{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward","family":"Darzynkiewicz","sequence":"additional","affiliation":[{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]},{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2511-7091","authenticated-orcid":true,"given":"Maciej","family":"Lukaszewicz","sequence":"additional","affiliation":[{"name":"University of Warsaw , , ,","place":["Warsaw, Poland, 02-093"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,5,6]]},"reference":[{"key":"2026090117284202000_cit1","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1080\/10409230490513991","article-title":"Messenger\nRNA turnover in eukaryotes:\npathways and enzymes","volume":"39","author":"Meyer","year":"2010","journal-title":"Crit. Rev. Biochem. Mol.\nBiol."},{"key":"2026090117284202000_cit2","doi-asserted-by":"publisher","first-page":"452","DOI":"10.1016\/j.bbagrm.2014.04.007","article-title":"Decapping Scavenger\n(DcpS) enzyme: Advances in its structure, activity and roles in the\ncap-dependent mRNA metabolism","volume":"1839","author":"Milac","year":"2014","journal-title":"Biochim. Biophys.\nActa"},{"key":"2026090117284202000_cit3","first-page":"1","article-title":"New insight\ninto decapping enzymes and selective mRNA\ndecay","volume":"8","author":"Grudzien-Nogalska","year":"2016","journal-title":"WIREs RNA"},{"key":"2026090117284202000_cit4","doi-asserted-by":"publisher","first-page":"3590","DOI":"10.1093\/nar\/gkh687","article-title":"The \u201cscavenger\u201d m7GpppX pyrophosphatase\nactivity of Dcs1 modulates nutrient-induced responses in yeast","volume":"32","author":"Malys","year":"2004","journal-title":"Nucl. Acids Res."},{"key":"2026090117284202000_cit5","doi-asserted-by":"publisher","first-page":"1609","DOI":"10.1261\/rna.7690504","article-title":"Nematode m7GpppG and m32,2,7GpppG\ndecapping: Activities in Ascaris\nembryos and characterization of C. elegans scavenger DcpS","volume":"10","author":"Cohen","year":"2004","journal-title":"RNA"},{"key":"2026090117284202000_cit6","doi-asserted-by":"publisher","first-page":"436","DOI":"10.1016\/j.tibs.2004.06.008","article-title":"Cap-tabolism","volume":"29","author":"Cougot","year":"2004","journal-title":"Trends Biochem.\nSci."},{"key":"2026090117284202000_cit7","doi-asserted-by":"publisher","first-page":"4699","DOI":"10.1093\/emboj\/cdf448","article-title":"The scavenger\nmRNA decapping enzyme DcpS is a member of the HIT family of pyrophosphatases","volume":"21","author":"Liu","year":"2002","journal-title":"EMBO J."},{"key":"2026090117284202000_cit8","doi-asserted-by":"publisher","first-page":"496","DOI":"10.1101\/gr.161034.113","article-title":"From single-cell\nto cell-pool transcriptomes:\nstochasticity in gene expression and RNA splicing","volume":"24","author":"Marinov","year":"2014","journal-title":"Genome Res."},{"key":"2026090117284202000_cit9","doi-asserted-by":"publisher","first-page":"5860","DOI":"10.1073\/pnas.092538799","article-title":"Precision and functional\nspecificity\nin mRNA decay","volume":"99","author":"Wang","year":"2002","journal-title":"Proc. Natl. Acad. Sci. U. S.\nA."},{"key":"2026090117284202000_cit10","doi-asserted-by":"publisher","first-page":"1863","DOI":"10.1101\/gr.1272403","article-title":"Decay rates of\nhuman mRNAs: correlation with functional characteristics and sequence\nattributes","volume":"13","author":"Yang","year":"2003","journal-title":"Genome Res."},{"key":"2026090117284202000_cit11","doi-asserted-by":"publisher","first-page":"12081","DOI":"10.1073\/pnas.1635192100","article-title":"DcpS can act in the 5\u2032-3\u2019\nmRNA decay pathway in addition to the 3\u2032-5\u2032 pathway","volume":"100","author":"van Dijk","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090117284202000_cit12","doi-asserted-by":"publisher","first-page":"370","DOI":"10.1016\/j.jmb.2006.08.015","article-title":"Dcs2, a novel stress-induced modulator of m7GpppX pyrophosphatase activity that locates to P bodies","volume":"363","author":"Malys","year":"2006","journal-title":"J. Mol. Biol."},{"key":"2026090117284202000_cit13","doi-asserted-by":"publisher","first-page":"8003","DOI":"10.1021\/bi300781g","article-title":"7-methylguanosine\ndiphosphate (m7GDP) is not hydrolyzed, but strongly bound\nby decapping scavenger (DcpS) enzymes and potently inhibits their\nactivity","volume":"51","author":"Wypijewska","year":"2012","journal-title":"Biochemistry"},{"key":"2026090117284202000_cit14","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1093\/nar\/gku1251","article-title":"Elimination of cap structure generated by mRNA decay\ninvolves the new scavenger mRNA decapping enzyme Aph1\/FHIT together\nwith DcpS","volume":"43","author":"Taverniti","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"2026090117284202000_cit15","doi-asserted-by":"publisher","first-page":"1132","DOI":"10.1261\/rna.1008208","article-title":"DcpS scavenger\ndecapping enzyme can modulate pre-mRNA splicing","volume":"14","author":"Shen","year":"2008","journal-title":"RNA"},{"key":"2026090117284202000_cit16","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/j.molcel.2013.02.023","article-title":"The decapping scavenger enzyme DCS-1 controls microRNA\nlevel in Caenorhabditis elegans","volume":"50","author":"Bosse","year":"2013","journal-title":"Mol. Cell"},{"key":"2026090117284202000_cit17","doi-asserted-by":"publisher","first-page":"3163","DOI":"10.1093\/hmg\/ddv067","article-title":"Loss of the scavenger mRNA decapping\nenzyme DCPS causes syndromic intellectual disability with neuromuscular\ndefects","volume":"24","author":"Ng","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"2026090117284202000_cit18","doi-asserted-by":"publisher","first-page":"3172","DOI":"10.1093\/hmg\/ddv069","article-title":"Mutations in DCPS and EDC3 in autosomal\nrecessive intellectual disability indicate a crucial role for mRNA\ndecapping in neurodevelopment","volume":"24","author":"Ahmed","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"2026090117284202000_cit19","doi-asserted-by":"publisher","first-page":"4084","DOI":"10.1093\/hmg\/ddt258","article-title":"The DcpS inhibitor RG3039 improves survival, function\nand motor unit pathologies in two SMA mouse models","volume":"22","author":"Gogliotti","year":"2013","journal-title":"Hum. Mol. Genet."},{"key":"2026090117284202000_cit20","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1016\/j.ccell.2018.01.012","article-title":"Genome-wide\nCRISPR-Cas9 Screen Identifies Leukemia-Specific Dependence\non a Pre-mRNA Metabolic Pathway Regulated by DCPS","volume":"33","author":"Yamauchi","year":"2018","journal-title":"Cancer Cell"},{"key":"2026090117284202000_cit21","doi-asserted-by":"publisher","first-page":"1349","DOI":"10.1080\/15257770701533818","article-title":"Affinity of\ndinucleotide cap analogs for human decapping scavenger (hDcpS)","volume":"26","author":"Darzynkiewicz","year":"2007","journal-title":"Nucleosides, Nucleotides and Nucleic Acids"},{"key":"2026090117284202000_cit22","doi-asserted-by":"publisher","first-page":"3003","DOI":"10.1111\/j.1742-4658.2010.07709.x","article-title":"Structural\nrequirements for Caenorhabditis elegans DcpS substrates\nbased on fluorescence and HPLC enzyme kinetic studies","volume":"277","author":"Wypijewska","year":"2010","journal-title":"FEBS J."},{"key":"2026090117284202000_cit23","doi-asserted-by":"publisher","first-page":"6508","DOI":"10.1111\/febs.12553","article-title":"Analysis of decapping scavenger cap\ncomplex using modified cap analogs reveals molecular determinants\nfor efficient cap binding","volume":"280","author":"Wypijewska del Nogal","year":"2013","journal-title":"FEBS J."},{"key":"2026090117284202000_cit24","doi-asserted-by":"publisher","first-page":"285217","DOI":"10.1088\/0953-8984\/19\/28\/285217","article-title":"Interaction of human decapping scavenger\nwith 5\u2019mRNA cap analogues: structural requirements for catalytic\nactivity","volume":"19","author":"Darzynkiewicz","year":"2007","journal-title":"J Phys Condens Matter"},{"key":"2026090117284202000_cit25","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/S1097-2765(04)00180-7","article-title":"Insight into the structure, mechanism,\nand regulation\nof scavenger mRNA decapping activity","volume":"14","author":"Gu","year":"2004","journal-title":"Mol. Cell"},{"key":"2026090117284202000_cit26","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1038\/nchembio.1866","article-title":"An excess\nof catalytically required motions inhibit the scavenger decapping\nenzyme","volume":"11","author":"Neu","year":"2015","journal-title":"Nat. Chem. Biol."},{"key":"2026090117284202000_cit27","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/S0076-6879(00)26058-8","article-title":"Purification of proteins using polyhistidine affinity\ntags","volume":"326","author":"Bornhorst","year":"2000","journal-title":"Methods Enzymol."},{"key":"2026090117284202000_cit28","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.pep.2005.12.002","article-title":"Current strategies\nfor the use of affinity tags and tag removal for the purification\nof recombinant proteins","volume":"48","author":"Arnau","year":"2006","journal-title":"Prot Expr Purif"},{"key":"2026090117284202000_cit29","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1023\/B:jsfg.0000031965.37625.0e","article-title":"His tag effect on solubility\nof human proteins produced\nin Escherichia coli: a comparison between four expression\nvectorss","volume":"5","author":"Woenstenek","year":"2004","journal-title":"J Struct Funct Genomics"},{"key":"2026090117284202000_cit30","doi-asserted-by":"publisher","first-page":"152","DOI":"10.1016\/j.pep.2004.10.017","article-title":"Attachment of a\nhistidine tag to the minimal zinc finger protein of the Aspergillus nidulans gene regulatory protein Area causes a conformational\nchange at the DNA-binding site","volume":"39","author":"Chant","year":"2005","journal-title":"Protein Expression\nPurif."},{"key":"2026090117284202000_cit31","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1016\/j.molcatb.2009.07.002","article-title":"The N-terminal His-tag\nand the recombinant process affects the biochemical properties of Staphylococcus aureus lipase produced in Escherichia coli","volume":"61","author":"Horchani","year":"2009","journal-title":"J. Mol. Catal. B:.Enzym."},{"key":"2026090117284202000_cit32","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.pep.2011.09.001","article-title":"Overexpression\nof (His)6-tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography","volume":"81","author":"Zakalskiy","year":"2012","journal-title":"Protein Expression Purif."},{"key":"2026090117284202000_cit33","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1107\/S0907444906052024","article-title":"His-tag impact on structure","volume":"63","author":"Carson","year":"2007","journal-title":"Acta Crystallogr., Sect. D: Biol.Crystallogr."},{"key":"2026090117284202000_cit34","doi-asserted-by":"publisher","first-page":"5588","DOI":"10.1021\/bi1005464","article-title":"Protein N-Terminal\nProcessing: Substrate Specificity of Escherichia coli and Human Methionine\nAminopeptidases","volume":"49","author":"Xiao","year":"2010","journal-title":"Biochemistry"},{"key":"2026090117284202000_cit35","doi-asserted-by":"publisher","first-page":"2336","DOI":"10.1074\/mcp.M600225-MCP200","article-title":"The proteomics of N-terminal methionine\ncleavage","volume":"5","author":"Frottin","year":"2006","journal-title":"Mol. Cell. Proteomics"},{"key":"2026090117284202000_cit36","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1006\/bbrc.1999.0770","article-title":"Mass spectrometric\ndetermination of a novel modification\nof the N-terminus of histidine-tagged proteins expressed in bacteria","volume":"259","author":"Yan","year":"1999","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2026090117284202000_cit37","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1006\/bbrc.1999.1304","article-title":"Identification of a gluconic acid\nderivative attached to the N-terminus of histidine-tagged proteins\nexpressed in bacteria","volume":"262","author":"Yan","year":"1999","journal-title":"Biochem. Biophys. Res.\nCommun."},{"key":"2026090117284202000_cit38","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1006\/abio.1998.2990","article-title":"Spontaneous\nalpha-N-6-phosphogluconoylation of a \u2033His tag\u2033 in Escherichia\ncoli: the cause of extra mass of 258 or 178 Da in fusion proteins","volume":"267","author":"Geoghegan","year":"1999","journal-title":"Anal. Biochem."},{"key":"2026090117284202000_cit39","doi-asserted-by":"publisher","first-page":"3003","DOI":"10.1111\/j.1742-4658.2010.07709.x","article-title":"Structural\nrequirements for Caenorhabditis elegans DcpS substrates based on fluorescence\nand HPLC enzyme kinetic studies","volume":"277","author":"Wypijewska","year":"2010","journal-title":"FEBS J."},{"key":"2026090117284202000_cit40","doi-asserted-by":"publisher","first-page":"16427","DOI":"10.1074\/jbc.M800341200","article-title":"Mechanistic\nand kinetic analysis of the DcpS scavenger decapping enzyme","volume":"283","author":"Liu","year":"2008","journal-title":"J. Biol. Chem."},{"key":"2026090117284202000_cit41","doi-asserted-by":"publisher","first-page":"1412","DOI":"10.1261\/rna.7660804","article-title":"Functional\nanalysis of mRNA scavenger decapping enzymes","volume":"10","author":"Liu","year":"2004","journal-title":"RNA"},{"key":"2026090117284202000_cit42","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1107\/S0907444901002918","article-title":"Post-translational\nmodification of the N-terminal His tag interferes with the crystallization\nof the wild-type and mutant SH3 domains from chicken src tyrosine\nkinase","volume":"57","author":"Kim","year":"2001","journal-title":"Acta Crystallogr., Sect. D: Biol.Crystallogr."},{"key":"2026090117284202000_cit43","doi-asserted-by":"publisher","first-page":"1359","DOI":"10.1002\/pro.2520","article-title":"Double trouble\n- Buffer selection and His-tag presence may be responsible for nonreproducibility\nof biomedical experiments","volume":"23","author":"Majorek","year":"2014","journal-title":"Protein Sci."},{"key":"2026090117284202000_cit44","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1081\/DMR-120033999","article-title":"Modeling kinetic data from in vitro drug metabolism\nenzyme experiments","volume":"36","author":"Tracy","year":"2004","journal-title":"Drug Metab. Rev."},{"key":"2026090117284202000_cit45","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1080\/15257779508012457","article-title":"Synthesis and properties of P1,P2-, P1,P3, and P1,P4-dinucleoside\ndi-, tri- and tetraphosphate mRNA cap analogues","volume":"14","author":"Stepinski","year":"1995","journal-title":"Nucleosides Nucleotides"},{"key":"2026090117284202000_cit46","doi-asserted-by":"crossref","first-page":"S197","DOI":"10.1135\/cccc1996s197","article-title":"1H NMR and fluorescence studies of new mRNA 5\u2032- cap analogues","volume":"61","author":"Jankowska","year":"1996","journal-title":"Coll. Czech. Chem. Commun."},{"key":"2026090117284202000_cit47","doi-asserted-by":"publisher","first-page":"17576","DOI":"10.1021\/acsomega.9b02715","article-title":"Decapping Scavenger Enzyme Activity toward N2-Substituted\n5\u2032 End mRNA Cap Analogues","volume":"4","author":"Pietrow","year":"2019","journal-title":"ACS Omega"},{"key":"2026090117284202000_cit48","doi-asserted-by":"publisher","first-page":"2212","DOI":"10.1038\/nprot.2007.321","article-title":"The use of differential\nscanning\nfluorimetry to detect ligand interactions that promote protein stability","volume":"2","author":"Niesen","year":"2007","journal-title":"Nat. Protoc."},{"key":"2026090117284202000_cit49","doi-asserted-by":"publisher","first-page":"167","DOI":"10.1016\/j.str.2016.11.018","article-title":"Interplay between Protein\nThermal Flexibility and Kinetic Stability","volume":"25","author":"Quezada","year":"2017","journal-title":"Structure"}],"container-title":["ACS Omega"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsomega.0c00304","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/5\/19\/10759\/67527651\/acsomega.0c00304.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/5\/19\/10759\/67527651\/acsomega.0c00304.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T21:28:48Z","timestamp":1788298128000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acsodf\/article\/5\/19\/10759\/615895\/Effect-of-the-His-Tag-Location-on-Decapping"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,6]]},"references-count":49,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2020,5,6]]},"published-print":{"date-parts":[[2020,5,19]]}},"URL":"https:\/\/doi.org\/10.1021\/acsomega.0c00304","relation":{},"ISSN":["2470-1343"],"issn-type":[{"value":"2470-1343","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,6]]}}}