The belonging of gpMuc, a glycoprotein from Mucuna pruriens seeds, to the Kunitz-type trypsin inhibitor family explains its direct anti-snake venom activity |
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Authors: | Andrea Scirè Fabio Tanfani,Enrico BertoliEmiliano Furlani,Hope-Onyekwere NNadozieHelena Cerutti,Alessio CortelazzoLuca Bini,Roberto Guerranti |
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Affiliation: | a Dipartimento di Biochimica, Biologia, e Genetica, Università Politecnica delle Marche, Via Ranieri, 60131 Ancona, Italy b Dipartimento di Medicina Interna, Scienze Endocrino-Metaboliche e Biochimica, Università di Siena, Via A. Moro, 53100 Siena, Italy c Dipartimento di Biologia Molecolare, Università di Siena, 53100 Siena, Italy |
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Abstract: | In Nigeria, Mucuna pruriens seeds are locally prescribed as an oral prophylactic for snake bite and it is claimed that when two seeds are swallowed they protect the individual for a year against snake bites. In order to understand the Mucuna pruriens antisnake properties, the proteins from the acqueous extract of seeds were purified by three chromatographic steps: ConA affinity chromatography, tandem anionic-cationic exchange and gel filtration, obtaining a fraction conventionally called gpMucB. This purified fraction was analysed by SDS-PAGE obtaining 3 bands with apparent masses ranging from 20 to 24 kDa, and by MALDI-TOF which showed two main peaks of 21 and 23 kDa and another small peak of 19 kDa. On the other hand, gel filtration analysis of the native protein indicated a molecular mass of about 70 kDa suggesting that in its native form, gpMucB is most likely an oligomeric multiform protein. Infrared spectroscopy of gpMucB indicated that the protein is particularly thermostable both at neutral and acidic pHs and that it is an all beta protein.All data suggest that gpMucB belongs to the Kunitz-type trypsin inhibitor family explaining the direct anti-snake venom activity of Mucuna pruriens seeds. |
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Keywords: | MPE, acqueous extract of Mucuna pruriens seeds gpMuc, glycoprotein from Mucuna pruriens seeds (ConA fraction) gpMucA, Ettan LC fraction gpMucB, EttanLC/gel filtration fraction FTIR, Fourier transform infrared Amide I&prime , amide I band in 2H2O medium K-PI, Kunitz-type trypsin inhibitor |
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