The Role of Primer in Glycogen Biosynthesis in Aerobacter aerogenes<sup>*</sup>

Tj, Kindt; He, Conrad

doi:10.1021/bi00864a014

Cited by 25 publications

(18 citation statements)

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“…The same ratio of components was maintained in reaction mixtures of larger volume. Reaction mixtures were incubated at 37 C, and the radioactivity incorporated into alcoholinsoluble polysaccharides was determined by spotting samples (50 Mliters) on filter paper squares, washing the papers in 66% ethanol, and measuring the retained radioactivity as described elsewhere (10,18). A unit of dextransucrase or levansucrase is arbitrarily defined as that amount of enzyme that incorporates 1 smol of glucose or fructose, respectively, from sucrose into polysaccharide per min at pH 6.8 and 37 C. Dextransucrase activities in Table 1 were determined from the amount of fructose released in the assay digests.…”

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confidence: 99%

Origin of the Cell-Associated Dextransucrase of Streptococcus mutans

McCabe

Smith

1973

Infect Immun

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The cell-associated dextransucrase produced by sucrose-grown cells of cariogenic Streptococcus mutans K1-R is derived from soluble dextransucrase. Synthesis of insoluble dextran by soluble dextransucrase gives rise to two dextransucrase fractions bound to the insoluble polysaccharide; a reversibly bound enzyme, which can be eluted in solutions of clinical dextran, and an irreversibly bound enzyme, which cannot be solubilized in this manner. Both of these fractions of dextransucrase are also present on sucrose-grown cells. During the synthesis of insoluble dextran by sucrose-grown cells, dextransucrase is progressively converted from soluble enzyme, first to the reversibly bound fraction and then to the irreversibly bound fraction, and is finally inactivated as insoluble dextran accumulates. The two cell-associated dextransucrase fractions therefore represent two stages in the insolubilization and inactivation of their precursor, soluble dextransucrase. As a result of this process of inactivation, the yield of dextransucrase from cells cultured on sucrose is markedly decreased by high concentrations of sucrose in the culture medium.

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confidence: 99%

Origin of the Cell-Associated Dextransucrase of Streptococcus mutans

McCabe

Smith

1973

Infect Immun

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“…U -14 C-Sucrose was purchased from New England Nuclear. The incorporation of U-14C-glucose into the total dextran was measured by placing aliquots of 14 C-sucrose-dextransucrase digests on squares of filter paper, and washing the paper with 66% ethanol and then acetone to remove all low molecular weight radioactive molecules, as by Kindt and Conrad [6]. The papers were dried and counted in scintillation solution containing 5 g diphenyloxazole and 0.5 g p-bis [2-(5-phenyloxazolyl)]-benzene per liter of toluene, using aPackard Tri-carb scintillation spectrometer.…”

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confidence: 99%

Biosynthetic relation between the soluble and insoluble dextrans produced by Leuconostoc mesenteroides NRRL B‐1299

Smith

1970

FEBS Letters

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“…Paper chromatograms were run in 1-butanol-pyridine-water (6:4:3) and standards were detected by spraying with aniline acid phthalate reagent (Partridge, 1949). Radioactive compounds on chromatograms were quantitated by cutting the chromatograms into segments and counting the segments as before (Kindt and Conrad, 1967).…”

Section: Methodsmentioning

confidence: 99%

“…The diversity of physical and chemical properties of the compounds which stimulate glycogen synthetase suggests that they are all activators of glycogen synthetase rather than acceptors for the glucosyl resic V_J"lycogen synthetases, regardless of their source, have shown an absolute requirement for a glycogentype glucan to accept glucose from the system-specific nucleoside diphosphate glucose (Leloir and Cardini, 1962;Greenberg and Preiss, 1965;Preiss and Greenberg, 1965). In previous work (Kindt and Conrad, 1967) it was shown that extracts of late log cells of Aerobacter aerogenes A3(S1) would synthesize glycogen from ADP-glucose only when exogenous glycogen was added, in spite of the fact that up to 2 g of glycogen/100 g of dry cells could be extracted from stationary-phase cultures of this organism. The tentative conclusion based on these observations was that late-log cells, in contrast to stationary-phase cells, were devoid of glycogen.…”

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confidence: 99%