DETECTION OF GLYCOGEN-DEBRANCHING SYSTEM IN TROPHOZOITES OF ENTAMOEBA-HISTOLYTICA

Autor(en): WERRIES, E
FRANZ, A
GEISEMEYER, S
Stichwörter: 4-ALPHA-GLUCANOTRANSFERASE; AMYLO-1,6-GLUCOSIDASE; BETA-AMYLASE; BINDING; BODIES; ENTAMOEBA-HISTOLYTICA; ENZYME; GLYCOGEN-DEBRANCHING SYSTEM; METABOLISM; MUSCLE; PHOSPHORYLASE; PURIFICATION; ULTRASTRUCTURE; Zoology
Erscheinungsdatum: 1990
Herausgeber: SOC PROTOZOOLOGISTS
Journal: JOURNAL OF PROTOZOOLOGY
Volumen: 37
Ausgabe: 6
Startseite: 576
Seitenende: 580
Zusammenfassung: 
Homogenates of trophozoites of Entamoeba histolytica were shown to bring about the total degradation of glycogen while purified phosphorylase of the same source alone yielded a limit dextrin as end product. An enzyme system capable of debranching the limit dextrin was obtained from the 40,000 g pellet by extraction in aqueous medium, purified by gel filtration on Fractogel TSK HW-55(F), and separated from phosphorylase by chromatography on Blue Sepharose CL-6B and aminobutyl Agarose. The glycogendebranching system was purified 540-fold to a state of homogeneity by criterion of disc-gel electrophoresis. The purified enzyme was able to degrade glycogen-limit dextrin in the presence of phosphorylase and exhibited activities of both amylo-1,6-glucosidase (EC 3.2.1.33) and 4-alpha-glucanotransferase (EC 2.4.1.25). Although amylo-1,6-glucosidase released glucose from a glycogen-phosphorylase limit dextrin, transferase activity moved single glucose residues from the limit dextrin to 4-nitrophenyl-alpha-glucoside yielding successively 4-nitrophenyl-alpha-maltoside and 4-nitrophenyl-alpha-maltotrioside that could be detected by HPLC. Native glycogen-debranching system exhibited a relative molecular mass of M(r) = 180,000 /- 10% by gel filtration and gel electrophoresis in both denaturing and nondenaturating conditions.
ISSN: 00223921
DOI: 10.1111/j.1550-7408.1990.tb01268.x

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