The Experts below are selected from a list of 159 Experts worldwide ranked by ideXlab platform
Victor Sanchezmargalet - One of the best experts on this subject based on the ideXlab platform.
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affinity purification of pancreastatin receptor gq 11 protein complex from rat liver membranes
Archives of Biochemistry and Biophysics, 2000Co-Authors: Jose Santosalvarez, Victor SanchezmargaletAbstract:Abstract Pancreastatin, a chromogranin A derived peptide, exerts a glycogenolytic effect on the hepatocyte. This effect is initiated by binding to membrane receptors which are coupled to pertussis toxin insensitive G proteins belonging to the Gq/11 family. We have recently solubilized active pancreastatin receptors from rat liver membranes still functionally coupled to G proteins. Here, we have purified pancreastatin receptors by a two-step procedure. First, pancreastatin receptors with their associated Gq/11 regulatory proteins were purified from liver membranes by lectin Absorption Chromatography on wheat germ agglutinin immobilized on agarose. A biotinylated rat pancreastatin analog was tested for binding to liver membranes before using it for affinity purification. Unlabeled biotinylated rat pancreastatin competed for 125 I-labeled [Tyr 0 ]PST binding to solubilized receptors with a K d = 0.27 nM, comparable to that of native pancreastatin. The biotinylated analog was immobilized on streptavidin-coated Sepharose beads and used to further affinity purify wheat germ agglutinin eluted receptor material. Specific elution at low pH showed that the receptor protein was purified as an 80-kDa protein in association with a G protein of the q/11 family, as demonstrated by specific immunoblot analysis. The specificity of the receptor band was assessed by chemical cross-linking of the purified material followed by SDS–PAGE and autoradiography. In conclusion, we have purified pancreastatin receptor as a glycoprotein of 80 kDa physically associated with a Gq/11 protein.
Víctor Sánchez-margalet - One of the best experts on this subject based on the ideXlab platform.
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Affinity purification of pancreastatin receptor-Gq/11 protein complex from rat liver membranes.
Archives of Biochemistry and Biophysics, 2000Co-Authors: José Santos-alvarez, Víctor Sánchez-margaletAbstract:Abstract Pancreastatin, a chromogranin A derived peptide, exerts a glycogenolytic effect on the hepatocyte. This effect is initiated by binding to membrane receptors which are coupled to pertussis toxin insensitive G proteins belonging to the Gq/11 family. We have recently solubilized active pancreastatin receptors from rat liver membranes still functionally coupled to G proteins. Here, we have purified pancreastatin receptors by a two-step procedure. First, pancreastatin receptors with their associated Gq/11 regulatory proteins were purified from liver membranes by lectin Absorption Chromatography on wheat germ agglutinin immobilized on agarose. A biotinylated rat pancreastatin analog was tested for binding to liver membranes before using it for affinity purification. Unlabeled biotinylated rat pancreastatin competed for 125 I-labeled [Tyr 0 ]PST binding to solubilized receptors with a K d = 0.27 nM, comparable to that of native pancreastatin. The biotinylated analog was immobilized on streptavidin-coated Sepharose beads and used to further affinity purify wheat germ agglutinin eluted receptor material. Specific elution at low pH showed that the receptor protein was purified as an 80-kDa protein in association with a G protein of the q/11 family, as demonstrated by specific immunoblot analysis. The specificity of the receptor band was assessed by chemical cross-linking of the purified material followed by SDS–PAGE and autoradiography. In conclusion, we have purified pancreastatin receptor as a glycoprotein of 80 kDa physically associated with a Gq/11 protein.
José Santos-alvarez - One of the best experts on this subject based on the ideXlab platform.
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Affinity purification of pancreastatin receptor-Gq/11 protein complex from rat liver membranes.
Archives of Biochemistry and Biophysics, 2000Co-Authors: José Santos-alvarez, Víctor Sánchez-margaletAbstract:Abstract Pancreastatin, a chromogranin A derived peptide, exerts a glycogenolytic effect on the hepatocyte. This effect is initiated by binding to membrane receptors which are coupled to pertussis toxin insensitive G proteins belonging to the Gq/11 family. We have recently solubilized active pancreastatin receptors from rat liver membranes still functionally coupled to G proteins. Here, we have purified pancreastatin receptors by a two-step procedure. First, pancreastatin receptors with their associated Gq/11 regulatory proteins were purified from liver membranes by lectin Absorption Chromatography on wheat germ agglutinin immobilized on agarose. A biotinylated rat pancreastatin analog was tested for binding to liver membranes before using it for affinity purification. Unlabeled biotinylated rat pancreastatin competed for 125 I-labeled [Tyr 0 ]PST binding to solubilized receptors with a K d = 0.27 nM, comparable to that of native pancreastatin. The biotinylated analog was immobilized on streptavidin-coated Sepharose beads and used to further affinity purify wheat germ agglutinin eluted receptor material. Specific elution at low pH showed that the receptor protein was purified as an 80-kDa protein in association with a G protein of the q/11 family, as demonstrated by specific immunoblot analysis. The specificity of the receptor band was assessed by chemical cross-linking of the purified material followed by SDS–PAGE and autoradiography. In conclusion, we have purified pancreastatin receptor as a glycoprotein of 80 kDa physically associated with a Gq/11 protein.
Jose Santosalvarez - One of the best experts on this subject based on the ideXlab platform.
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affinity purification of pancreastatin receptor gq 11 protein complex from rat liver membranes
Archives of Biochemistry and Biophysics, 2000Co-Authors: Jose Santosalvarez, Victor SanchezmargaletAbstract:Abstract Pancreastatin, a chromogranin A derived peptide, exerts a glycogenolytic effect on the hepatocyte. This effect is initiated by binding to membrane receptors which are coupled to pertussis toxin insensitive G proteins belonging to the Gq/11 family. We have recently solubilized active pancreastatin receptors from rat liver membranes still functionally coupled to G proteins. Here, we have purified pancreastatin receptors by a two-step procedure. First, pancreastatin receptors with their associated Gq/11 regulatory proteins were purified from liver membranes by lectin Absorption Chromatography on wheat germ agglutinin immobilized on agarose. A biotinylated rat pancreastatin analog was tested for binding to liver membranes before using it for affinity purification. Unlabeled biotinylated rat pancreastatin competed for 125 I-labeled [Tyr 0 ]PST binding to solubilized receptors with a K d = 0.27 nM, comparable to that of native pancreastatin. The biotinylated analog was immobilized on streptavidin-coated Sepharose beads and used to further affinity purify wheat germ agglutinin eluted receptor material. Specific elution at low pH showed that the receptor protein was purified as an 80-kDa protein in association with a G protein of the q/11 family, as demonstrated by specific immunoblot analysis. The specificity of the receptor band was assessed by chemical cross-linking of the purified material followed by SDS–PAGE and autoradiography. In conclusion, we have purified pancreastatin receptor as a glycoprotein of 80 kDa physically associated with a Gq/11 protein.
Lidija Petrushevska‐tozi - One of the best experts on this subject based on the ideXlab platform.
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Mineral and water soluble vitamin content in the Kombucha drink
International Journal of Food Science and Technology, 2000Co-Authors: Biljana Bauer‐petrovska, Lidija Petrushevska‐toziAbstract:Biologically active substances (water soluble vitamins and minerals) were analyzed in 'Kombucha' a curative liquor, produced by the so-called 'Kombucha cultivation' (Macedonian collection of microorganisms, No 734) in sweet black tea decoct. The water soluble vitamins in the Kombucha drink were separated and identified using thin layer Chromatography (TLC) and quantified by the comparison of their chromatographic UV spectra with the reference compounds. Four soluble vitamins have been determinated to have the following concentrations: vitamin B 1 0.74 mg ml -1 , vitamin B 6 0.52 mg ml -1 , vitamin B 12 0.84 mg ml -1 and vitamin C 1.51 mg ml -1 . Mineral elements of nutritional and toxicological importance were determined in dissolved ash using atomic Absorption Chromatography (AAS). Mineral composition content involved determination of the essential elements: zinc, copper, iron, manganese, nickel and cobalt. Investigations of some toxic elements showed that lead and chromium were present in very small amounts, whereas cadmium was not found.
