LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies
Both ricin and <i>R. communis</i> <i>agglutinin</i> (RCA120), belonging to the type II ribosome-inactivating proteins (RIPs-Ⅱ), are derived from the seeds of the castor bean plant. They share very similar amino acid sequences, but ricin is much more toxic than RCA120. It is u...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-07-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/11/7/393 |
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| author | Long-Hui Liang Chang-Cai Liu Bo Chen Long Yan Hui-Lan Yu Yang Yang Ji-Na Wu Xiao-Sen Li Shi-Lei Liu |
| author_facet | Long-Hui Liang Chang-Cai Liu Bo Chen Long Yan Hui-Lan Yu Yang Yang Ji-Na Wu Xiao-Sen Li Shi-Lei Liu |
| author_sort | Long-Hui Liang |
| collection | DOAJ |
| description | Both ricin and <i>R. communis</i> <i>agglutinin</i> (RCA120), belonging to the type II ribosome-inactivating proteins (RIPs-Ⅱ), are derived from the seeds of the castor bean plant. They share very similar amino acid sequences, but ricin is much more toxic than RCA120. It is urgently necessary to distinguish ricin and RCA120 in response to public safety. Currently, mass spectrometric assays are well established for unambiguous identification of ricin by accurate analysis of differentiated amino acid residues after trypsin digestion. However, diagnostic peptides are relatively limited for unambiguous identification of trace ricin, especially in complex matrices. Here, we demonstrate a digestion strategy of multiple proteinases to produce novel peptide markers for unambiguous identification of ricin. Liquid chromatography-high resolution MS (LC-HRMS) was used to verify the resulting peptides, among which only the peptides with uniqueness and good MS response were selected as peptide markers. Seven novel peptide markers were obtained from tandem digestion of trypsin and endoproteinase Glu-C in PBS buffer. From the chymotrypsin digestion under reduction and non-reduction conditions, eight and seven novel peptides were selected respectively. Using pepsin under pH 1~2 and proteinase K digestion, six and five peptides were selected as novel peptide markers. In conclusion, the obtained novel peptides from the established digestion methods can be recommended for the unambiguous identification of ricin during the investigation of illegal use of the toxin. |
| first_indexed | 2024-04-11T22:28:22Z |
| format | Article |
| id | doaj.art-514422dafc4e40eab9d61d15f1537f9b |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T22:28:22Z |
| publishDate | 2019-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-514422dafc4e40eab9d61d15f1537f9b2022-12-22T03:59:36ZengMDPI AGToxins2072-66512019-07-0111739310.3390/toxins11070393toxins11070393LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion StrategiesLong-Hui Liang0Chang-Cai Liu1Bo Chen2Long Yan3Hui-Lan Yu4Yang Yang5Ji-Na Wu6Xiao-Sen Li7Shi-Lei Liu8State Key Laboratory of NBC Protection for Civilian, Beijing 102205, ChinaState Key Laboratory of NBC Protection for Civilian, Beijing 102205, ChinaThe laboratory of Analytical Chemistry, Research Institute of Chemical Defence, Beijing 102205, ChinaThe laboratory of Analytical Chemistry, Research Institute of Chemical Defence, Beijing 102205, ChinaState Key Laboratory of NBC Protection for Civilian, Beijing 102205, ChinaState Key Laboratory of NBC Protection for Civilian, Beijing 102205, ChinaThe laboratory of Analytical Chemistry, Research Institute of Chemical Defence, Beijing 102205, ChinaThe laboratory of Analytical Chemistry, Research Institute of Chemical Defence, Beijing 102205, ChinaState Key Laboratory of NBC Protection for Civilian, Beijing 102205, ChinaBoth ricin and <i>R. communis</i> <i>agglutinin</i> (RCA120), belonging to the type II ribosome-inactivating proteins (RIPs-Ⅱ), are derived from the seeds of the castor bean plant. They share very similar amino acid sequences, but ricin is much more toxic than RCA120. It is urgently necessary to distinguish ricin and RCA120 in response to public safety. Currently, mass spectrometric assays are well established for unambiguous identification of ricin by accurate analysis of differentiated amino acid residues after trypsin digestion. However, diagnostic peptides are relatively limited for unambiguous identification of trace ricin, especially in complex matrices. Here, we demonstrate a digestion strategy of multiple proteinases to produce novel peptide markers for unambiguous identification of ricin. Liquid chromatography-high resolution MS (LC-HRMS) was used to verify the resulting peptides, among which only the peptides with uniqueness and good MS response were selected as peptide markers. Seven novel peptide markers were obtained from tandem digestion of trypsin and endoproteinase Glu-C in PBS buffer. From the chymotrypsin digestion under reduction and non-reduction conditions, eight and seven novel peptides were selected respectively. Using pepsin under pH 1~2 and proteinase K digestion, six and five peptides were selected as novel peptide markers. In conclusion, the obtained novel peptides from the established digestion methods can be recommended for the unambiguous identification of ricin during the investigation of illegal use of the toxin.https://www.mdpi.com/2072-6651/11/7/393ricinmarker peptidesunambiguous identificationmass spectrometry |
| spellingShingle | Long-Hui Liang Chang-Cai Liu Bo Chen Long Yan Hui-Lan Yu Yang Yang Ji-Na Wu Xiao-Sen Li Shi-Lei Liu LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies Toxins ricin marker peptides unambiguous identification mass spectrometry |
| title | LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies |
| title_full | LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies |
| title_fullStr | LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies |
| title_full_unstemmed | LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies |
| title_short | LC-HRMS Screening and Identification of Novel Peptide Markers of Ricin Based on Multiple Protease Digestion Strategies |
| title_sort | lc hrms screening and identification of novel peptide markers of ricin based on multiple protease digestion strategies |
| topic | ricin marker peptides unambiguous identification mass spectrometry |
| url | https://www.mdpi.com/2072-6651/11/7/393 |
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