Analysis of the free nucleotide pools of mammalian tissues by high-pressure liquid chromatography

Brown, Eric G.; Newton, Russell P.; Shaw, Nicholas M.

doi:10.1016/0003-2697(82)90461-4

Cited by 74 publications

(20 citation statements)

References 14 publications

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“…. Furthermore, Honeyfield et al (2005) experimentally reproduced early mortality syndrome in lake trout fry and observed adult mortality (as previously reported by Brown et al [2005b]). Signs and symptoms of EMS were observed when fish were fed diets containing thiaminase (Honeyfield et al, 2005).…”

Section: Discussionsupporting

confidence: 74%

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Pathology, Physiologic Parameters, Tissue Contaminants, and Tissue Thiamine in Morbid and Healthy Central Florida Adult American Alligators (Alligator Mississippiensis)

Honeyfield

Ross

Carbonneau

et al. 2008

Journal of Wildlife Diseases

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ABSTRACT:An investigation of adult alligator (Alligator mississippiensis) mortalities in Lake Griffin, central Florida, was conducted from 1998-2004. Alligator mortality was highest in the months of April and May and annual death count peaked in 2000. Bacterial pathogens, heavy metals, and pesticides were not linked with the mortalities. Blood chemistry did not point to any clinical diagnosis, although differences between impaired and normal animals were noted. Captured alligators with signs of neurologic impairment displayed unresponsive and uncoordinated behavior. Three of 21 impaired Lake Griffin alligators were found to have neural lesions characteristic of thiamine deficiency in the telencephalon, particularly the dorsal ventricular ridge. In some cases, lesions were found in the thalamus, and parts of the midbrain. Liver and muscle tissue concentrations of thiamine (vitamin B 1 ) were lowest in impaired Lake Griffin alligators when compared to unimpaired alligators or to alligators from Lake Woodruff. The consumption of thiaminase-positive gizzard shad (Dorosoma cepedianum) is thought to have been the cause of the low tissue thiamine and resulting mortalities.

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Section: Discussionsupporting

confidence: 74%

“…Analyses were completed as described in Brown et al (1998) with minor modifications of the elution gradient. All reagents were made with Burdick Jackson HPLC grade water (VWR Scientific Products, S. Plainfield, New Jersey, USA).…”

Section: Thiamine Analysismentioning

confidence: 99%

Pathology, Physiologic Parameters, Tissue Contaminants, and Tissue Thiamine in Morbid and Healthy Central Florida Adult American Alligators (Alligator Mississippiensis)

Honeyfield

Ross

Carbonneau

et al. 2008

Journal of Wildlife Diseases

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“…The resolution of a reference mixture of uridine, UMP, UDP-Glc, UDPGal, and UDP wasˆrst checked. Under the elution conditions of Brown et al, 11) the sugar nucleotides were not separated from one another. The elution gradient was modiˆed to theˆrst 30 min isocratic, and the next 30 min gradient elution.…”

Section: Resultsmentioning

confidence: 99%

“…According to Brown et al, 11) 5 mM KH2PO4 (pH 3) and 500 mM (pH 4) were used as the initial eluent (BuŠer A) and as theˆnal (BuŠer B) respectively. In theˆrst 30 min, 10z of BuŠer B was run isocratically, followed by a linear gradient of 10-100z B in the next 30 min.…”

Section: Chemicals Udp-d-[u-mentioning

confidence: 99%

Production of Galactinol from Sucrose by Plant Enzymes

Wakiuchi

Shiomi

Tamaki

2003

Bioscience, Biotechnology, and Biochemistry

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Galactinol, 1-O-(a-D-galactopyranosyl)-myo-inositol, was produced from sucrose as a starting material.UDP-Glc was prepared with sucrose and UDP using sucrose synthase partially puriˆed from sweet potato roots. Then, the UDP-Glc was converted to UDP-Gal using yeast UDP-Gal 4-epimerase from a commercial source. Finally, galactinol was produced from the UDPGal and myo-inositol using galactinol synthase partially puriˆed from cucumber leaves. The product was identied as galactinol by the retention times of HPLC, agalactosidase digestion, and NMR spectrometry.

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“…[␤-32 P]UDPGlcNAc was purified by reverse-phase high pressure liquid chromatography on an Ultrasphere ODS column (0.5 ϫ 30 cm) with a linear gradient from 40 mM KH 2 PO 4 , pH 6.0, 5 mM tetrabutylammonium dihydrogen phosphate to 100% acetonitrile at 1 ml/min (17). Alternatively, [␤-32 P]UDP-GlcNAc was purified by ion-exchange chromatography on a Microsorb MV column (0.25 ϫ 25 cm) equilibrated with 5 mM KH 2 PO 4 , pH 4.0, and developed at 1 ml/min with a linear gradient from 5 mM KH 2 PO 4 , pH 4.0, to 1.5 M KH 2 PO 4 , pH 3.0, as described (18). Purified [␤- 32 P]UDP-GlcNAc had a specific activity of 10 mCi/mol and was homogeneous in both chromatography systems.…”

Section: Glcnac-phosphotransferase Assaymentioning

confidence: 99%

Bovine UDP-N-acetylglucosamine:Lysosomal-enzyme N-Acetylglucosamine-1-phosphotransferase I

Bao

Booth

Elmendorf

et al. 1996

Journal of Biological Chemistry

123

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UDP-N-acetylglucosamine:lysosomal-enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-phosphotransferase) catalyzes the initial step in the synthesis of the mannose 6-phosphate determinant required for efficient intracellular targeting of newly synthesized lysosomal hydrolases to the lysosome. The enzyme was partially purified ϳ30,000-fold by chromatography of solubilized membrane proteins from lactating bovine mammary glands on DEAE-Sepharose, reactive green 19-agarose, and Superose 6. The partially purified enzyme was used to generate a panel of murine monoclonal antibodies. The antiGlcNAc-phosphotransferase monoclonal antibody PT18 was coupled to a solid support and used to immunopurify the enzyme ϳ480,000-fold to apparent homogeneity with an overall yield of 29%. The purified enzyme has a specific activity of 10-12 mol of GlcNAc phosphate transferred per h/mg using 100 mM ␣-methylmannoside as acceptor.The subunit structure of the enzyme was determined using a combination of analytical gel filtration chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and amino-terminal sequencing. The data indicate that bovine GlcNAc-phosphotransferase is a 540,000-Da complex composed of disulfide-linked homodimers of 166,000-and 51,000-Da subunits and two identical, noncovalently associated 56,000-Da subunits.The trafficking of most lysosomal hydrolases in higher eucaryotes is mediated by a mannose 6-phosphate-dependent pathway. Asparagine-linked oligosaccharides on newly synthesized lysosomal hydrolases are sequentially modified by two enzymes to generate a mannose 6-phosphate recognition marker. The initial and determining step in the biosynthesis of the mannose 6-phosphate recognition marker is catalyzed by the enzyme UDP-N-acetylglucosamine:lysosomal-enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-phosphotransferase).

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Analysis of the free nucleotide pools of mammalian tissues by high-pressure liquid chromatography

Cited by 74 publications

References 14 publications

Pathology, Physiologic Parameters, Tissue Contaminants, and Tissue Thiamine in Morbid and Healthy Central Florida Adult American Alligators (Alligator Mississippiensis)

Pathology, Physiologic Parameters, Tissue Contaminants, and Tissue Thiamine in Morbid and Healthy Central Florida Adult American Alligators (Alligator Mississippiensis)

Production of Galactinol from Sucrose by Plant Enzymes

Bovine UDP-N-acetylglucosamine:Lysosomal-enzyme N-Acetylglucosamine-1-phosphotransferase I

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