TY  - JOUR
A1  - Bieber, Elke
A1  - Woenckhaus, Christoph
A1  - Pauli, Harald
T1  - Preparation of an affinity chromatographic system for the separation of ADP binding proteins
T2  - Zeitschrift für Naturforschung, C
N2  - [4-(3-Bromoacetylpyridinio)-butyl]adenosine pyrophosphate as a structural analog of NAD+ reacts covalently with the sulfhydryl groups of thiopropyl agarose. 10-20 μmol can be bound to 1 ml gel. Stabilization of the insoluble coenzym e is attained by treatment with sodium boro hydride (NaBH4). This complex when applied to column chromatography, allow s the separation of various dehydrogenases as a result of their different complex stability coefficients. Alcohol dehydrogenase from liver, lactate dehydrogenase, and adenylate kinase, which all bind to the ADP-analog residues of the gel matrix, can thus be separated by different salt gradients. Alcohol dehydrogenase from yeast, however, does not form a complex and can easily be eluted from the column with phosphate buffer. Glyceraldehyde-3 phosphate and aldehyde dehydrogenases can be eluted by the addition of NAD+ or NADH to the buffer. The uncharged 1,4-dihydropyridin ring of the reduced coenzyme produces a more stable complex with the dehydrogenases than the oxidized form.
KW  - Affinity Chromatography
KW  - NAD+-Analogs
KW  - Separation of ADP-Binding Enzymes
Y1  - 2014
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/81787
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-817879
SN  - 0939-5075
SN  - 1865-7125
VL  - 39.1984
IS  - 11-12
SP  - 1041
EP  - 1047
PB  - Verlag der Zeitschrift für Naturforschung
CY  - Tübingen
ER  -