{"title":"评估亚硝基钴胺素与内在因子的结合亲和力,作为钴胺素结合蛋白相互作用的预测模型:与羟钴胺的比较研究。","authors":"Annette M Sysel, Joseph A Bauer","doi":"10.31083/FBE26810","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Intrinsic factor (IF) is a glycoprotein crucial for cobalamin (vitamin B<sub>12</sub>) absorption in the human body. This study aimed to evaluate the binding affinity of nitrosylcobalamin (NO-Cbl), a cobalamin analog, to recombinant human IF derived from plants, using hydroxocobalamin (OH-Cbl) as a comparative standard.</p><p><strong>Methods: </strong>Surface plasmon resonance (SPR) was employed to assess the kinetic parameters of NO-Cbl and OH-Cbl interactions with plant- derived IF across various concentrations.</p><p><strong>Results: </strong>SPR analysis demonstrated that NO-Cbl and OH-Cbl exhibited high binding affinities to IF, with equilibrium dissociation constant (<i>K</i><sub>D</sub>) values in the picomolar range. OH-Cbl showed a slightly stronger binding affinity (<i>K</i><sub>D</sub> = 4.79 × 10<sup>-11</sup> M) than NO-Cbl (<i>K</i><sub>D</sub> = 8.58 × 10<sup>-11</sup> M). Despite NO-Cbl and OH-Cbl both being bound to IF, differences in binding affinity and stability were observed, particularly at higher concentrations.</p><p><strong>Conclusion: </strong>Variations in IF binding between NO-Cbl and OH-Cbl may be attributed to the saturation of binding sites or recognition issues specific to plant-derived IF. This study underscores the potential of NO-Cbl as a targeted therapeutic agent capable of leveraging natural cobalamin uptake pathways. These results also highlight the suitability of using recombinant plant-derived IF as a model for predicting the biological activity of cobalamin analogs despite the nuanced differences from native human IF.</p>","PeriodicalId":73068,"journal":{"name":"Frontiers in bioscience (Elite edition)","volume":"17 1","pages":"26810"},"PeriodicalIF":0.0000,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Evaluation of the Binding Affinity of Nitrosylcobalamin to Intrinsic Factor as a Predictive Model for Cobalamin Binding Protein Interactions: A Comparative Study with Hydroxocobalamin.\",\"authors\":\"Annette M Sysel, Joseph A Bauer\",\"doi\":\"10.31083/FBE26810\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Intrinsic factor (IF) is a glycoprotein crucial for cobalamin (vitamin B<sub>12</sub>) absorption in the human body. This study aimed to evaluate the binding affinity of nitrosylcobalamin (NO-Cbl), a cobalamin analog, to recombinant human IF derived from plants, using hydroxocobalamin (OH-Cbl) as a comparative standard.</p><p><strong>Methods: </strong>Surface plasmon resonance (SPR) was employed to assess the kinetic parameters of NO-Cbl and OH-Cbl interactions with plant- derived IF across various concentrations.</p><p><strong>Results: </strong>SPR analysis demonstrated that NO-Cbl and OH-Cbl exhibited high binding affinities to IF, with equilibrium dissociation constant (<i>K</i><sub>D</sub>) values in the picomolar range. OH-Cbl showed a slightly stronger binding affinity (<i>K</i><sub>D</sub> = 4.79 × 10<sup>-11</sup> M) than NO-Cbl (<i>K</i><sub>D</sub> = 8.58 × 10<sup>-11</sup> M). Despite NO-Cbl and OH-Cbl both being bound to IF, differences in binding affinity and stability were observed, particularly at higher concentrations.</p><p><strong>Conclusion: </strong>Variations in IF binding between NO-Cbl and OH-Cbl may be attributed to the saturation of binding sites or recognition issues specific to plant-derived IF. This study underscores the potential of NO-Cbl as a targeted therapeutic agent capable of leveraging natural cobalamin uptake pathways. These results also highlight the suitability of using recombinant plant-derived IF as a model for predicting the biological activity of cobalamin analogs despite the nuanced differences from native human IF.</p>\",\"PeriodicalId\":73068,\"journal\":{\"name\":\"Frontiers in bioscience (Elite edition)\",\"volume\":\"17 1\",\"pages\":\"26810\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-03-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Frontiers in bioscience (Elite edition)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.31083/FBE26810\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in bioscience (Elite edition)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.31083/FBE26810","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Evaluation of the Binding Affinity of Nitrosylcobalamin to Intrinsic Factor as a Predictive Model for Cobalamin Binding Protein Interactions: A Comparative Study with Hydroxocobalamin.
Background: Intrinsic factor (IF) is a glycoprotein crucial for cobalamin (vitamin B12) absorption in the human body. This study aimed to evaluate the binding affinity of nitrosylcobalamin (NO-Cbl), a cobalamin analog, to recombinant human IF derived from plants, using hydroxocobalamin (OH-Cbl) as a comparative standard.
Methods: Surface plasmon resonance (SPR) was employed to assess the kinetic parameters of NO-Cbl and OH-Cbl interactions with plant- derived IF across various concentrations.
Results: SPR analysis demonstrated that NO-Cbl and OH-Cbl exhibited high binding affinities to IF, with equilibrium dissociation constant (KD) values in the picomolar range. OH-Cbl showed a slightly stronger binding affinity (KD = 4.79 × 10-11 M) than NO-Cbl (KD = 8.58 × 10-11 M). Despite NO-Cbl and OH-Cbl both being bound to IF, differences in binding affinity and stability were observed, particularly at higher concentrations.
Conclusion: Variations in IF binding between NO-Cbl and OH-Cbl may be attributed to the saturation of binding sites or recognition issues specific to plant-derived IF. This study underscores the potential of NO-Cbl as a targeted therapeutic agent capable of leveraging natural cobalamin uptake pathways. These results also highlight the suitability of using recombinant plant-derived IF as a model for predicting the biological activity of cobalamin analogs despite the nuanced differences from native human IF.