A toxic thionin from Pyrularia pubera: purification, properties, and amino acid sequence |
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| Authors: | L P Vernon G E Evett R D Zeikus W R Gray |
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| Affiliation: | 1. Department of Chemistry, Brigham Young University, Provo, Utah 84602 USA;2. Department of Microbiology, Brigham Young University, Provo, Utah 84602, USA;3. Biology Department, University of Utah, Salt Lake City, Utah 84112 USA;1. Sharda University, Material Research Laboratory, School of Basic Sciences and Research, Greater Noida 201310, U.P., India;2. Department of Micro and Nano System Technology (IMST), HBV-Buskerud & Vestfold University College, Raveien 197, Horten, 3103 Vestfold, Norway;3. Sharda University, Department of Electronics and Communication, Greater Noida 201310, U.P., India;1. Vavilov Institute of General Genetics, Russian Academy of Sciences, 3 Gubkina Str., 119991 Moscow, Russian Federation;2. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya Str., 117997 Moscow, Russian Federation;3. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilova Str., Moscow 119991, Russian Federation;1. Faculty of Environment and Labour Safety, Ton Duc Thang University, Ho Chi Minh City, Vietnam;2. Faculty of Environmental Sciences, University of Science, Vietnam National University, 334 Nguyen Trai, Thanh Xuan, Hanoi, Vietnam;3. NTT Hi-Tech Institute, Nguyen Tat Thanh University, Ho Chi Minh City 700000, Vietnam;4. Faculty of Chemistry, University of Science, Vietnam National University, 334 Nguyen Trai, Thanh Xuan, Hanoi, Vietnam;5. VNU Key Laboratory of Advanced Materials for Green Growth, University of Science, Vietnam National University, 334 Nguyen Trai, Thanh Xuan, Hanoi, Vietnam;1. Department of Chemistry, KTH Royal Institute of Technology, 10044 Stockholm, Sweden, Sweden;2. Department of Physics and Astronomy, Uppsala University, 751 20 Uppsala, Sweden;3. State Key Laboratory of Fine Chemicals, DUT-KTH Joint Education and Research Center on Molecular Devices, Dalian University of Technology (DUT), Dalian 116024, PR China |
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| Abstract: | A low-molecular-weight cytotoxic protein has been purified from Pyrularia pubera Michx. (Santalaceae). By comparison with the behavior of proteins of known molecular weight during Sephadex G-75 gel filtration and denaturing electrophoresis, a molecular weight of somewhat less than 6000 is indicated. Purification involves ammonium sulfate fractionation followed by either gel filtration on Sephadex G-75 or separation on a carboxymethyl cellulose CM52 column. At concentrations of 0.04 mg/ml the protein causes visible disruption of cultured mouse B16 melanoma cells. The complete amino acid sequence has been determined. The toxin contains 47 amino acids arranged as follows:Lys-Ser-Cys-Cys-Arg-Asn-Thr-Trp-Ala-Arg-Asn-C ys-Tyr-Asn-Val-Cys-Arg-Leu-Pro-Gly-Thr-Ile-Ser-Arg-Glu-Ile-Cys-Ala-Lys- Lys-Cys-Asp-Cys-Lys-Ile-Ile-Ser-Gly-Thr-Thr-Cys-Pro-Ser-Asp-Tyr-Pro-Ly s-OH. The protein is clearly a thionin, as shown by its close resemblance to the thionins from wheat and barley, to the viscotoxins from mistletoes, and to crambin. |
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