Isolation and characterization of Glyceraldehyde-3-phosphate dehydrogenase from the common octopus (Octopus vulgaris Cuvier, 1797)

Isolation and characterization of Glyceraldehyde-3-phosphate dehydrogenase from the common... The NAD+ dependent cytosolic Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) from arms of Octopus vulgaris, Cuvier, 1787, (Octopoda, Cephalopoda) was purified to homogeneity and its kinetic properties investigated. The purification method consisted of ammonium sulfate fractionation followed by Blue Sepharose CL-6B chromatography resulting in a 26-fold increase in specific activity and a final yield of approximately 16%. The apparent molecular weight of the purified native enzyme was 153 kDa. The protein is an homotetramer, composed of identical subunits with an apparent molecular weight of approximately 36 kDa. The Michaelis constants Km for both NAD+ and d-G3P were 66 μM and 320 μM, respectively. The maximal velocity Vmax of the purified enzyme was estimated to be 21.8 U/mg. Only one GAPDH isoform (pI 6.6) was obtained by isoelectrofocusing in polyacrylamide slab gels holding ampholyte generated pH gradients. Under the conditions of assay, the optimum activity occurs at pH 7.0 and at temperature of 35°C. Polyclonal antibodies raised in rabbits against the purified GAPDH immunostained a single 36 kDa GAPDH band on crude extract protein preparations blotted onto nitrocellulose. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Reviews in Fish Biology and Fisheries Springer Journals

Isolation and characterization of Glyceraldehyde-3-phosphate dehydrogenase from the common octopus (Octopus vulgaris Cuvier, 1797)

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Publisher
Springer Netherlands
Copyright
Copyright © 2007 by Springer Science+Business Media B.V.
Subject
Life Sciences; Zoology ; Freshwater & Marine Ecology
ISSN
0960-3166
eISSN
1573-5184
D.O.I.
10.1007/s11160-007-9074-6
Publisher site
See Article on Publisher Site

References

  • Cephalopod and groundfish landings: evidence for ecological change in global fisheries?
    Caddy, JF; Rodhouse, PG

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