{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:35:44Z","timestamp":1760146544349,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,11,16]],"date-time":"2024-11-16T00:00:00Z","timestamp":1731715200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"CRIS projects","award":["2030-32000-011-000-D","5030-32000-228-000-D"],"award-info":[{"award-number":["2030-32000-011-000-D","5030-32000-228-000-D"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Chronic wasting disease (CWD) is a prion disease afflicting wild and farmed elk. CWD prions (PrPSc) are infectious protein conformations that replicate by inducing a natively expressed prion protein (PrPC) to refold into the prion conformation. Mass spectrometry was used to study the prions resulting from a previously described experimental inoculation of MM132, ML132, and LL132 elk with a common CWD inoculum. Chymotryptic digestion times and instrument parameters were optimized to yield a set of six peptides, TNMK, MLGSAMSRPL, LLGSAMSRPL, ENMYR, MMER, and VVEQMCITQYQR. These peptides were used to quantify the amount, the M132 and L132 polymorphic composition, and the extent of methionine oxidation of elk PrPSc. The amount (ng\/g brain tissue) of PrPSc present in each sample was determined to be: MM132 (5.4 \u00d7 102 \u00b1 7 \u00d7 101), ML132 (3.3 \u00d7 102 \u00b1 6 \u00d7 101 and 3.6 \u00d7 102 \u00b1 3 \u00d7 101) and LL132 (0.7 \u00d7 102 \u00b1 1 \u00d7 101, 0.2 \u00d7 102 \u00b1 0.2 \u00d7 101, and 0.2 \u00d7 102 \u00b1 0.5 \u00d7 101). The proportion of L132 polymorphism in ML132 (heterozygous) PrPSc from CWD-infected elk was determined to be 43% \u00b1 2% or 36% \u00b1 3%. Methionine oxidation was detected and quantified for the M132 and L132 polymorphisms in the samples. In this way, mass spectrometry can be used to characterize prion strains at a molecular level.<\/jats:p>","DOI":"10.3390\/pathogens13111008","type":"journal-article","created":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T04:07:09Z","timestamp":1731902829000},"page":"1008","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Quantifying the Molecular Properties of the Elk Chronic Wasting Disease Agent with Mass Spectrometry"],"prefix":"10.3390","volume":"13","author":[{"given":"Christopher J.","family":"Silva","sequence":"first","affiliation":[{"name":"Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Albany, CA 94710, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0505-9568","authenticated-orcid":false,"given":"Melissa L.","family":"Erickson-Beltran","sequence":"additional","affiliation":[{"name":"Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Albany, CA 94710, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7130-2650","authenticated-orcid":false,"given":"Eric D.","family":"Cassmann","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Patterson Hall, 1800 Christensen Drive, Ames, IA 50011, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2202-3054","authenticated-orcid":false,"given":"Justin J.","family":"Greenlee","sequence":"additional","affiliation":[{"name":"Virus and Prion Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA 50010, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"89","DOI":"10.7589\/0090-3558-16.1.89","article-title":"Chronic wasting disease of captive mule deer: A spongiform encephalopathy","volume":"16","author":"Williams","year":"1980","journal-title":"J. Wildl. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.3201\/eid1006.040010","article-title":"Environmental sources of prion transmission in mule deer","volume":"10","author":"Miller","year":"2004","journal-title":"Emerg. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1038\/425035a","article-title":"Prion disease: Horizontal prion transmission in mule deer","volume":"425","author":"Miller","year":"2003","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"465","DOI":"10.7589\/0090-3558-18.4.465","article-title":"Spongiform encephalopathy of Rocky Mountain elk","volume":"18","author":"Williams","year":"1982","journal-title":"J. Wildl. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1292\/jvms.64.855","article-title":"A case of chronic wasting disease in an elk imported to Korea from Canada","volume":"64","author":"Sohn","year":"2002","journal-title":"J. Vet. Med. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"S15","DOI":"10.1080\/19336896.2016.1163048","article-title":"Prion 2016 Animal Prion Disease Workshop Abstracts","volume":"10","author":"Sohn","year":"2016","journal-title":"Prion"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1292\/jvms.67.753","article-title":"Additional cases of Chronic Wasting Disease in imported deer in Korea","volume":"67","author":"Kim","year":"2005","journal-title":"J. Vet. Med. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1186\/s13567-016-0375-4","article-title":"First case of chronic wasting disease in Europe in a Norwegian free-ranging reindeer","volume":"47","author":"Benestad","year":"2016","journal-title":"Vet. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"31417","DOI":"10.1073\/pnas.2013237117","article-title":"Studies in bank voles reveal strain differences between chronic wasting disease prions from Norway and North America","volume":"117","author":"Nonno","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"461","DOI":"10.7589\/JWD-D-20-00141","article-title":"First Detection of Chronic Wasting Disease in Moose (Alces alces) in Sweden","volume":"57","author":"Agren","year":"2021","journal-title":"J. Wildl. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.3201\/eid2412.180702","article-title":"Novel Type of Chronic Wasting Disease Detected in Moose (Alces alces), Norway","volume":"24","author":"Pirisinu","year":"2018","journal-title":"Emerg. Infect. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"970","DOI":"10.7589\/2018-10-262","article-title":"First Detection of Chronic Wasting Disease in a Wild Red Deer (Cervus elaphus) in Europe","volume":"55","author":"Vikoren","year":"2019","journal-title":"J. Wildl. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"323","DOI":"10.3201\/eid2902.220882","article-title":"Novel Prion Strain as Cause of Chronic Wasting Disease in a Moose, Finland","volume":"29","author":"Sun","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1186\/s13567-024-01320-y","article-title":"Classical BSE dismissed as the cause of CWD in Norwegian red deer despite strain similarities between both prion agents","volume":"55","author":"Benestad","year":"2024","journal-title":"Vet. Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Hopp, P., Rolandsen, C.M., Korpenfelt, S.L., Vage, J., Soren, K., Solberg, E.J., Averhed, G., Pusenius, J., Rosendal, T., and Ericsson, G. (2024). Sporadic cases of chronic wasting disease in old moose\u2014An epidemiological study. J. Gen. Virol., 105.","DOI":"10.1099\/jgv.0.001952"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Arifin, M.I., Hannaoui, S., Ng, R.A., Zeng, D., Zemlyankina, I., Ahmed-Hassan, H., Schatzl, H.M., Kaczmarczyk, L., Jackson, W.S., and Benestad, S.L. (2024). Norwegian moose CWD induces clinical disease and neuroinvasion in gene-targeted mice expressing cervid S138N prion protein. PLoS Pathog., 20.","DOI":"10.1371\/journal.ppat.1012350"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1126\/science.6801762","article-title":"Novel proteinaceous infectious particles cause scrapie","volume":"216","author":"Prusiner","year":"1982","journal-title":"Science"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"13363","DOI":"10.1073\/pnas.95.23.13363","article-title":"Prions","volume":"95","author":"Prusiner","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","unstructured":"Gajdusek, C.J., Gibbs, C.J., and Alpers, M.P. (1965). Experiments with Scrapie with Special Reference to the Nature of the Agent and the Pathology of the Disease, Slow, Latent and Temperate Virus Infections (NINDB Monograph 2)."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2711","DOI":"10.1099\/0022-1317-68-10-2711","article-title":"Linkage of the scrapie-associated fibril protein (PrP) gene and Sinc using congenic mice and restriction fragment length polymorphism analysis","volume":"68","author":"Hunter","year":"1987","journal-title":"J. Gen. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Arifin, M.I., Hannaoui, S., Chang, S.C., Thapa, S., Schatzl, H.M., and Gilch, S. (2021). Cervid Prion Protein Polymorphisms: Role in Chronic Wasting Disease Pathogenesis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052271"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2765","DOI":"10.1099\/0022-1317-80-10-2765","article-title":"PrP genotypes of captive and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni) with chronic wasting disease","volume":"80","author":"Besser","year":"1999","journal-title":"J. Gen. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1177\/104063870601800118","article-title":"Preliminary observations of genetic susceptibility of elk (Cervus elaphus nelsoni) to chronic wasting disease by experimental oral inoculation","volume":"18","author":"Hamir","year":"2006","journal-title":"J. Vet. Diagn. Invest."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1935","DOI":"10.1097\/WNR.0b013e3282f1ca2f","article-title":"Elk with a long incubation prion disease phenotype have a unique PrPd profile","volume":"18","author":"Spraker","year":"2007","journal-title":"Neuroreport"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Moore, S.J., Vrentas, C.E., Hwang, S., West Greenlee, M.H., Nicholson, E.M., and Greenlee, J.J. (2018). Pathologic and biochemical characterization of PrP(Sc) from elk with PRNP polymorphisms at codon 132 after experimental infection with the chronic wasting disease agent. BMC Vet. Res., 14.","DOI":"10.1186\/s12917-018-1400-9"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Moore, J., Tatum, T., Hwang, S., Vrentas, C., West Greenlee, M.H., Kong, Q., Nicholson, E., and Greenlee, J. (2020). Novel Strain of the Chronic Wasting Disease Agent Isolated from Experimentally Inoculated Elk with LL132 Prion Protein. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-59819-1"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1070","DOI":"10.1016\/j.jasms.2007.03.009","article-title":"Mass spectrometric detection of attomole amounts of the prion protein by nanoLC\/MS\/MS","volume":"18","author":"Onisko","year":"2007","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1021\/acs.analchem.6b03822","article-title":"Quantitating PrP Polymorphisms Present in Prions from Heterozygous Scrapie-Infected Sheep","volume":"89","author":"Silva","year":"2017","journal-title":"Anal. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1021\/acs.analchem.7b03985","article-title":"Determining the Relative Susceptibility of Four Prion Protein Genotypes to Atypical Scrapie","volume":"90","author":"Silva","year":"2018","journal-title":"Anal. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1276","DOI":"10.1021\/acs.analchem.9b04449","article-title":"A General Mass Spectrometry-Based Method of Quantitating Prion Polymorphisms from Heterozygous Chronic Wasting Disease-Infected Cervids","volume":"92","author":"Silva","year":"2020","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1021\/jasms.2c00280","article-title":"General Method of Quantifying the Extent of Methionine Oxidation in the Prion Protein","volume":"34","author":"Silva","year":"2023","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2905","DOI":"10.1099\/0022-1317-72-12-2905","article-title":"Copurification of Sp33\u201337 and scrapie agent from hamster brain prior to detectable histopathology and clinical disease","volume":"72","author":"Bolton","year":"1991","journal-title":"J. Gen. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1021\/ac102527w","article-title":"Utility of mass spectrometry in the diagnosis of prion diseases","volume":"83","author":"Silva","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2793","DOI":"10.1073\/pnas.90.7.2793","article-title":"Attempts to restore scrapie prion infectivity after exposure to protein denaturants","volume":"90","author":"Prusiner","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2139","DOI":"10.1021\/bi3016795","article-title":"Oxidation of methionine 216 in sheep and elk prion protein is highly dependent upon the amino acid at position 218 but is not important for prion propagation","volume":"52","author":"Silva","year":"2013","journal-title":"Biochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Lourenc\u0327o Dos Santos, S., Petropoulos, I., and Friguet, B. (2018). The Oxidized Protein Repair Enzymes Methionine Sulfoxide Reductases and Their Roles in Protecting against Oxidative Stress, in Ageing and in Regulating Protein Function. Antioxidants, 7.","DOI":"10.3390\/antiox7120191"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1007\/s11064-017-2460-0","article-title":"Methionine in Proteins: It\u2019s Not Just for Protein Initiation Anymore","volume":"44","author":"Lim","year":"2019","journal-title":"Neurochem. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1854","DOI":"10.1021\/bi901850n","article-title":"Assessing the role of oxidized methionine at position 213 in the formation of prions in hamsters","volume":"49","author":"Silva","year":"2010","journal-title":"Biochemistry"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2279","DOI":"10.1021\/acs.jafc.0c06241","article-title":"Time of Detection of Prions in the Brain by Nanoscale Liquid Chromatography Coupled to Tandem Mass Spectrometry Is Comparable to Animal Bioassay","volume":"69","author":"Silva","year":"2021","journal-title":"J. Agric. Food Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1073\/pnas.0409008102","article-title":"Prion protein NMR structures of elk and of mouse\/elk hybrids","volume":"102","author":"Gossert","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/11\/1008\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:33:33Z","timestamp":1760114013000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/13\/11\/1008"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,16]]},"references-count":40,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["pathogens13111008"],"URL":"https:\/\/doi.org\/10.3390\/pathogens13111008","relation":{},"ISSN":["2076-0817"],"issn-type":[{"type":"electronic","value":"2076-0817"}],"subject":[],"published":{"date-parts":[[2024,11,16]]}}}