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A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes

Standard

A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes. / Hagel, C; Fleissner, A; Seifert, R.

In: ANAL BIOCHEM, Vol. 182, No. 1, 01.10.1989, p. 64-70.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Hagel, C, Fleissner, A & Seifert, R 1989, 'A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes', ANAL BIOCHEM, vol. 182, no. 1, pp. 64-70.

APA

Hagel, C., Fleissner, A., & Seifert, R. (1989). A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes. ANAL BIOCHEM, 182(1), 64-70.

Vancouver

Hagel C, Fleissner A, Seifert R. A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes. ANAL BIOCHEM. 1989 Oct 1;182(1):64-70.

Bibtex

@article{87675bc9c680420bbbbf6987a9a01e01,
title = "A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes",
abstract = "A radiochemical microassay for soluble and membrane-bound glutamate decarboxylase (GAD) is described. Up to 180 samples can be determined per day with a variation coefficient of 2%. The method detects newly synthesized gamma-amino-n-butyric acid in the picomole range and can easily be applied to other enzymes whose substrate and product differ by charge. In an aqueous homogenate of brain (1 + 10; w/v) about 15% of the total GAD activity are spun down by centrifugation (1 h, 100,000g) increasing to 35% of the total GAD activity in solutions with 8 mM calcium chloride or 100 mM potassium acetate. There is similar dependence on the cation concentration when GAD binds to phospholipid vesicles (liposomes) as well as dependence on lipid concentration and lipid composition. The coenzyme pyridoxal 5'-phosphate has no influence on GAD binding to liposomes.",
keywords = "Animals, Carbon Radioisotopes, Glutamate Decarboxylase, Glutamates, Glutamic Acid, Liposomes, Male, Membrane Proteins, Microchemistry, Protein Binding, Rats, Solubility, gamma-Aminobutyric Acid",
author = "C Hagel and A Fleissner and R Seifert",
year = "1989",
month = oct,
day = "1",
language = "English",
volume = "182",
pages = "64--70",
journal = "ANAL BIOCHEM",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

RIS

TY - JOUR

T1 - A microassay for the determination of soluble and membrane-bound glutamate decarboxylase activity--influences of cations, lipid composition, and pyridoxal 5'-phosphate on the glutamate decarboxylase binding to liposomes

AU - Hagel, C

AU - Fleissner, A

AU - Seifert, R

PY - 1989/10/1

Y1 - 1989/10/1

N2 - A radiochemical microassay for soluble and membrane-bound glutamate decarboxylase (GAD) is described. Up to 180 samples can be determined per day with a variation coefficient of 2%. The method detects newly synthesized gamma-amino-n-butyric acid in the picomole range and can easily be applied to other enzymes whose substrate and product differ by charge. In an aqueous homogenate of brain (1 + 10; w/v) about 15% of the total GAD activity are spun down by centrifugation (1 h, 100,000g) increasing to 35% of the total GAD activity in solutions with 8 mM calcium chloride or 100 mM potassium acetate. There is similar dependence on the cation concentration when GAD binds to phospholipid vesicles (liposomes) as well as dependence on lipid concentration and lipid composition. The coenzyme pyridoxal 5'-phosphate has no influence on GAD binding to liposomes.

AB - A radiochemical microassay for soluble and membrane-bound glutamate decarboxylase (GAD) is described. Up to 180 samples can be determined per day with a variation coefficient of 2%. The method detects newly synthesized gamma-amino-n-butyric acid in the picomole range and can easily be applied to other enzymes whose substrate and product differ by charge. In an aqueous homogenate of brain (1 + 10; w/v) about 15% of the total GAD activity are spun down by centrifugation (1 h, 100,000g) increasing to 35% of the total GAD activity in solutions with 8 mM calcium chloride or 100 mM potassium acetate. There is similar dependence on the cation concentration when GAD binds to phospholipid vesicles (liposomes) as well as dependence on lipid concentration and lipid composition. The coenzyme pyridoxal 5'-phosphate has no influence on GAD binding to liposomes.

KW - Animals

KW - Carbon Radioisotopes

KW - Glutamate Decarboxylase

KW - Glutamates

KW - Glutamic Acid

KW - Liposomes

KW - Male

KW - Membrane Proteins

KW - Microchemistry

KW - Protein Binding

KW - Rats

KW - Solubility

KW - gamma-Aminobutyric Acid

M3 - SCORING: Journal article

C2 - 2574957

VL - 182

SP - 64

EP - 70

JO - ANAL BIOCHEM

JF - ANAL BIOCHEM

SN - 0003-2697

IS - 1

ER -