A comparison of two different analytical workflows to determine the venom proteome composition of Naja kaouthia from North-East India and immunological profiling of venom against commercial antivenoms
The Indian monocled cobra (Naja kaouthia) is one of the most prevalent venomous snakes in northeast India (NEI) and is the cause of many fatalities. The composition of NEI N. kaouthia venom (NkV) was deciphered using two different proteomic approaches: (i) 1D SDS-PAGE coupled to label-free quantific...
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| Veröffentlicht in: | International journal of biological macromolecules 2022-05, Vol.208, p.275-287 |
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| creator | Kakati, Hirakjyoti Patra, Aparup Kalita, Bhargab Chanda, Abhishek Rapole, Srikanth Mukherjee, Ashis K. |
| description | The Indian monocled cobra (Naja kaouthia) is one of the most prevalent venomous snakes in northeast India (NEI) and is the cause of many fatalities. The composition of NEI N. kaouthia venom (NkV) was deciphered using two different proteomic approaches: (i) 1D SDS-PAGE coupled to label-free quantification of protein bands using stringent identification criteria and (ii) reversed-phase high-performance liquid chromatography (RP-HPLC) followed by quantification based on area under the RP-HPLC peaks. The proteomic data from both strategies were compared. Proteomic analyses from both workflows identified 32 proteins (toxins) distributed over 10–14 snake venom protein families in NEI NkV. The relative abundances of the venom proteins determined from the analytical workflows coincided with the densitometry band intensities of the NEI NkV. Phospholipase A2 (13.1–16.0%) and three-finger toxins (58.5–64.2%) represented the most abundant enzymatic and non-enzymatic proteins in NEI NkV, respectively. Immuno-cross-reactivity studies by enzyme-linked immunoassay and immunoblot analyses pointed to the poor efficacy of commercial PAVs in recognizing the low molecular mass ( |
| doi_str_mv | 10.1016/j.ijbiomac.2022.03.095 |
| format | Article |
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•Each method of venom de-complexation for protein identification and quantification has its pros and cons•The venom proteome composition of N. kaouthia from North-East India was deciphered using two proteomic workflows and compared•The study demonstrated that both analytical workflows can be considered reliable approaches for analyzing snake venom proteome•The 3FTx and PLA2 represented the most abundant non-enzymatic and enzymatic proteins in the tested venom•Immunoprofiling demonstrated poor efficacy of commercial PAVs against the low molecular weight (<15 kDa) NEI NKVtoxins</description><identifier>ISSN: 0141-8130</identifier><identifier>EISSN: 1879-0003</identifier><identifier>DOI: 10.1016/j.ijbiomac.2022.03.095</identifier><identifier>PMID: 35331793</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Indian monocled cobra ; LC-MS/MS analysis ; Venom proteomics ; Venom-antivenom interaction</subject><ispartof>International journal of biological macromolecules, 2022-05, Vol.208, p.275-287</ispartof><rights>2022 Elsevier B.V.</rights><rights>Copyright © 2022 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c368t-d591263500f7b498f0eae319304ec024ad3785e3519fe9c431865e0fba233b433</citedby><cites>FETCH-LOGICAL-c368t-d591263500f7b498f0eae319304ec024ad3785e3519fe9c431865e0fba233b433</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ijbiomac.2022.03.095$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35331793$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kakati, Hirakjyoti</creatorcontrib><creatorcontrib>Patra, Aparup</creatorcontrib><creatorcontrib>Kalita, Bhargab</creatorcontrib><creatorcontrib>Chanda, Abhishek</creatorcontrib><creatorcontrib>Rapole, Srikanth</creatorcontrib><creatorcontrib>Mukherjee, Ashis K.</creatorcontrib><title>A comparison of two different analytical workflows to determine the venom proteome composition of Naja kaouthia from North-East India and immunological profiling of venom against commercial antivenoms</title><title>International journal of biological macromolecules</title><addtitle>Int J Biol Macromol</addtitle><description>The Indian monocled cobra (Naja kaouthia) is one of the most prevalent venomous snakes in northeast India (NEI) and is the cause of many fatalities. The composition of NEI N. kaouthia venom (NkV) was deciphered using two different proteomic approaches: (i) 1D SDS-PAGE coupled to label-free quantification of protein bands using stringent identification criteria and (ii) reversed-phase high-performance liquid chromatography (RP-HPLC) followed by quantification based on area under the RP-HPLC peaks. The proteomic data from both strategies were compared. Proteomic analyses from both workflows identified 32 proteins (toxins) distributed over 10–14 snake venom protein families in NEI NkV. The relative abundances of the venom proteins determined from the analytical workflows coincided with the densitometry band intensities of the NEI NkV. Phospholipase A2 (13.1–16.0%) and three-finger toxins (58.5–64.2%) represented the most abundant enzymatic and non-enzymatic proteins in NEI NkV, respectively. Immuno-cross-reactivity studies by enzyme-linked immunoassay and immunoblot analyses pointed to the poor efficacy of commercial PAVs in recognizing the low molecular mass (<15 kDa) toxins of NEI NkV. Spectrofluorometric titration determined the presence of NEI NkV-specific antibodies in commercial PAV, at a level that was higher than that previously reported for eastern India NkV-specific antibodies in commercial antivenom.
•Each method of venom de-complexation for protein identification and quantification has its pros and cons•The venom proteome composition of N. kaouthia from North-East India was deciphered using two proteomic workflows and compared•The study demonstrated that both analytical workflows can be considered reliable approaches for analyzing snake venom proteome•The 3FTx and PLA2 represented the most abundant non-enzymatic and enzymatic proteins in the tested venom•Immunoprofiling demonstrated poor efficacy of commercial PAVs against the low molecular weight (<15 kDa) NEI NKVtoxins</description><subject>Indian monocled cobra</subject><subject>LC-MS/MS analysis</subject><subject>Venom proteomics</subject><subject>Venom-antivenom interaction</subject><issn>0141-8130</issn><issn>1879-0003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNqFkcFu3CAURVHVqpmk_YWIZTeegLE99q5RlLSRonTTrhHGj5nnGJgCzih_2M8qM0667Qrpce69Dy4hl5ytOePN1bjGsUdvlV6XrCzXTKxZV78jK95uuoIxJt6TFeMVL1ou2Bk5j3HM06bm7UdyJmoh-KYTK_Lnmmpv9ypg9I56Q9PB0wGNgQAuUeXU9JJQq4kefHgykz9EmjIBCYJFBzTtgD6D85bug0_gLZwMfcSEi-OjGhV9Un5OO1TUhIw--pB2xa2Kid67IU-VGyhaOzs_-e0pLrsZnNBtjxZLgNoqdFmS_S0EjZlSLuHpMn4iH4yaInx-PS_Ir7vbnzffi4cf3-5vrh8KLZo2FUPd8bIRNWNm01ddaxgoELwTrALNykoNYtPWIGreGeh0JXjb1MBMr0oh-kqIC_Jl8c0L_p4hJmkxapgm5cDPUZZNVeV_F3Wb0WZBdfAxBjByH9Cq8CI5k8cW5SjfWpTHFiUTMreYhZevGXNvYfgne6stA18XAPJLnxGCjBrBaRgwgE5y8Pi_jL9vj7av</recordid><startdate>20220531</startdate><enddate>20220531</enddate><creator>Kakati, Hirakjyoti</creator><creator>Patra, Aparup</creator><creator>Kalita, Bhargab</creator><creator>Chanda, Abhishek</creator><creator>Rapole, Srikanth</creator><creator>Mukherjee, Ashis K.</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20220531</creationdate><title>A comparison of two different analytical workflows to determine the venom proteome composition of Naja kaouthia from North-East India and immunological profiling of venom against commercial antivenoms</title><author>Kakati, Hirakjyoti ; Patra, Aparup ; Kalita, Bhargab ; Chanda, Abhishek ; Rapole, Srikanth ; Mukherjee, Ashis K.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c368t-d591263500f7b498f0eae319304ec024ad3785e3519fe9c431865e0fba233b433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Indian monocled cobra</topic><topic>LC-MS/MS analysis</topic><topic>Venom proteomics</topic><topic>Venom-antivenom interaction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kakati, Hirakjyoti</creatorcontrib><creatorcontrib>Patra, Aparup</creatorcontrib><creatorcontrib>Kalita, Bhargab</creatorcontrib><creatorcontrib>Chanda, Abhishek</creatorcontrib><creatorcontrib>Rapole, Srikanth</creatorcontrib><creatorcontrib>Mukherjee, Ashis K.</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>International journal of biological macromolecules</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kakati, Hirakjyoti</au><au>Patra, Aparup</au><au>Kalita, Bhargab</au><au>Chanda, Abhishek</au><au>Rapole, Srikanth</au><au>Mukherjee, Ashis K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A comparison of two different analytical workflows to determine the venom proteome composition of Naja kaouthia from North-East India and immunological profiling of venom against commercial antivenoms</atitle><jtitle>International journal of biological macromolecules</jtitle><addtitle>Int J Biol Macromol</addtitle><date>2022-05-31</date><risdate>2022</risdate><volume>208</volume><spage>275</spage><epage>287</epage><pages>275-287</pages><issn>0141-8130</issn><eissn>1879-0003</eissn><abstract>The Indian monocled cobra (Naja kaouthia) is one of the most prevalent venomous snakes in northeast India (NEI) and is the cause of many fatalities. The composition of NEI N. kaouthia venom (NkV) was deciphered using two different proteomic approaches: (i) 1D SDS-PAGE coupled to label-free quantification of protein bands using stringent identification criteria and (ii) reversed-phase high-performance liquid chromatography (RP-HPLC) followed by quantification based on area under the RP-HPLC peaks. The proteomic data from both strategies were compared. Proteomic analyses from both workflows identified 32 proteins (toxins) distributed over 10–14 snake venom protein families in NEI NkV. The relative abundances of the venom proteins determined from the analytical workflows coincided with the densitometry band intensities of the NEI NkV. Phospholipase A2 (13.1–16.0%) and three-finger toxins (58.5–64.2%) represented the most abundant enzymatic and non-enzymatic proteins in NEI NkV, respectively. Immuno-cross-reactivity studies by enzyme-linked immunoassay and immunoblot analyses pointed to the poor efficacy of commercial PAVs in recognizing the low molecular mass (<15 kDa) toxins of NEI NkV. Spectrofluorometric titration determined the presence of NEI NkV-specific antibodies in commercial PAV, at a level that was higher than that previously reported for eastern India NkV-specific antibodies in commercial antivenom.
•Each method of venom de-complexation for protein identification and quantification has its pros and cons•The venom proteome composition of N. kaouthia from North-East India was deciphered using two proteomic workflows and compared•The study demonstrated that both analytical workflows can be considered reliable approaches for analyzing snake venom proteome•The 3FTx and PLA2 represented the most abundant non-enzymatic and enzymatic proteins in the tested venom•Immunoprofiling demonstrated poor efficacy of commercial PAVs against the low molecular weight (<15 kDa) NEI NKVtoxins</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>35331793</pmid><doi>10.1016/j.ijbiomac.2022.03.095</doi><tpages>13</tpages></addata></record> |
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| subjects | Indian monocled cobra LC-MS/MS analysis Venom proteomics Venom-antivenom interaction |
| title | A comparison of two different analytical workflows to determine the venom proteome composition of Naja kaouthia from North-East India and immunological profiling of venom against commercial antivenoms |
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