A study of nucleotides and nucleotide sugars in human blood cells has been carried out. Ten samples from healthy yound people of group "O" were analyzed by ion-exchange chromatography, and the following nucleotides and nucleotide sugars were found: NAD+; UDP-glucose, UDP-mannose, UDP-galactose, UDP-acetylglucosamine and UDP-acetylgalactosamine; AMP; NADP+; ADP-glucose, ADP-galactose and ADP-mannose; GDP-mannose and GDP-fucose; ADP and ATP. The sugar nucleotides were identified on the basis of chemical, enzymatic and chromatographic analysis. UDP-galactose, UDP-mannose, UDP-acetylgalactosamine, ADP-glucose, ADP-galactose, ADP-mannose, GDP-mannose and GDP-fucose have not been found before in human blood, and UDP-mannose was not known to occur in nature. In order to minimize the decomposition of unstable substances, mild techniques were applied: cold acid extraction and neutral (ethanol-chloroform) extraction were compared, and in both cases the aqueous phase was passed through a column of Dowex-i (Cl-form) resin, and the nucleotides eluted with a linear gradient of NH4Cl. Results showed that the extraction step is not very critical, but the use of ammonium salts and almost neutral pH, as compared with methods used in previous papers, permitted the isolation of ten nucleotide-linked sugars, eight of which have not been described before in human blood. An adenine nucleotide with very particular properties was extracted with the ethanol-chloroform technique; it was eluted by the concentration of NH4Cl necessary to elute ATP, but it behaved as AMP on paper chromatography in several solvent systems. It is very unstable and decomposes even at -20° in a few days. © 1971.
Documento: | Artículo |
Título: | Nucleotides and nucleotide sugars in human blood cells. II. Group "O" |
Autor: | Cantore, M.L.; Leoni, P.; Leveroni, A.F.; Recondo, E.F. |
Filiación: | Instituto de Investigaciones Bioquímicas Fundacion Campomar, Argentina Facultad de Ciencias Exactas y Naturales, Obligado 2490 Buenos Aires (28), Argentina Ministerio de Bienestar Social de la Provincia de Buenos Aires Argentina |
Palabras clave: | adenine nucleotide; alcohol; ammonium chloride; carbon; chloroform; fucose; galactose; glucosamine; guanine nucleotide; hexosamine; hexose; mannose; nicotinamide adenine dinucleotide; nicotinamide adenine dinucleotide phosphate; nucleoside diphosphate sugar; nucleotide; pyrimidine nucleotide; trichloroacetic acid; adult; article; blood; blood cell; blood group ABO system; electrophoresis; glucose blood level; human; ion exchange chromatography; isolation and purification; paper; paper chromatography; pH; spectrophotometry; technique; ultraviolet radiation; ABO Blood-Group System; Adenine Nucleotides; Adult; Ammonium Chloride; Blood Cells; Blood Glucose; Carbon Isotopes; Chloroform; Chromatography, Ion Exchange; Chromatography, Paper; Electrophoresis; Ethanol; Fucose; Galactose; Glucosamine; Guanine Nucleotides; Hexosamines; Hexoses; Human; Hydrogen-Ion Concentration; Mannose; Methods; NAD; NADP; Nucleoside Diphosphate Sugars; Nucleotides; Paper; Spectrophotometry; Trichloroacetic Acid; Ultraviolet Rays; Uracil Nucleotides |
Año: | 1971 |
Volumen: | 230 |
Número: | 3 |
Página de inicio: | 423 |
Página de fin: | 433 |
DOI: | http://dx.doi.org/10.1016/0304-4165(71)90170-X |
Título revista: | BBA - General Subjects |
Título revista abreviado: | Biochim. Biophys. Acta Gen. Subj. |
ISSN: | 03044165 |
CODEN: | BBGSB |
CAS: | alcohol, 64-17-5; ammonium chloride, 12125-02-9; carbon, 7440-44-0; chloroform, 67-66-3; fucose, 3615-37-0, 3713-31-3; galactose, 26566-61-0, 50855-33-9, 59-23-4; glucosamine, 3416-24-8, 4607-22-1; hexose, 93780-23-5; mannose, 31103-86-3, 3458-28-4; nicotinamide adenine dinucleotide phosphate, 53-59-8; nicotinamide adenine dinucleotide, 53-84-9; trichloroacetic acid, 14357-05-2, 76-03-9; ABO Blood-Group System; Adenine Nucleotides; Ammonium Chloride, 12125-02-9; Blood Glucose; Carbon Isotopes; Chloroform, 67-66-3; Ethanol, 64-17-5; Fucose, 3713-31-3; Galactose, 26566-61-0; Glucosamine, 3416-24-8; Guanine Nucleotides; Hexosamines; Hexoses; Mannose, 31103-86-3; NAD, 53-84-9; NADP, 53-59-8; Nucleoside Diphosphate Sugars; Nucleotides; Trichloroacetic Acid, 76-03-9; Uracil Nucleotides |
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v230_n3_p423_Cantore |