Specificity and function of glyceraldehyde-3-phosphate dehydrogenase in a freshwater diatom, Asterionella formosa (Bacillariophyceae)

Erales, Jenny; Gontero, Brigitte; Maberly, Stephen C. ORCID: https://orcid.org/0000-0003-3541-5903. 2008 Specificity and function of glyceraldehyde-3-phosphate dehydrogenase in a freshwater diatom, Asterionella formosa (Bacillariophyceae). Journal of Phycology, 44. 1455-1464. 10.1111/j.1529-8817.2008.00600.x

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Abstract/Summary

The plastidic GAPDH catalyses the only reductive step in the Calvin cycle and exists as different forms of which GapC1 enzyme is present in chromalveolates such as diatoms. Biochemical studies on diatoms are still fragmentary and thus, in this report, GAPDH from the freshwater diatom Asterionella formosa Hassall, has been purified and kinetically characterised. It is a homotetrameric enzyme with a molecular mass of about 150 ± 15 kDa. The enzyme showed Michaelis-Menten kinetics with respect to both cofactors, NADPH and NADH, with a 16-fold greater catalytic constant for NADPH. The Km for NADPH was 140 µM, the lowest affinity reported, while the catalytic constant, 815 s-1, is the highest reported. The Km for NADH was 93 µM and the catalytic constant was 50 s-1, both are similar to reported values for other types of GAPDH. The GapC1 enzyme, like the Chlamydomonas reinhardtii P.A. Dang A4 GAPDH, exhibits a cooperative behaviour towards the substrate, BPGA, with both cofactors. Mass spectrometry analysis showed that when GapC1 enzyme was purified without reducing agents, it co-purified with a small protein with a mass of 8.2 kDa. This protein was recognized by antibodies against CP12. When associated with this protein, GAPDH displayed a lag that disappeared upon incubation with reducing agent in the presence of either BPGA or NADPH as a consequence of dissociation of the GAPDH/CP12 complex. Thus, as in other species of algae and higher plants, regulation of GapC1 enzyme in A. formosa may occur through association-dissociation processes linked to dark-light transitions.

Item Type:	Publication - Article
Digital Object Identifier (DOI):	10.1111/j.1529-8817.2008.00600.x
Programmes:	CEH Programmes pre-2009 publications > Water > WA02 Quantifying processes that link water quality and quantity, biota and physical environment > WA02.3 Physico-chemical processes and effects on freshwater biot
UKCEH and CEH Sections/Science Areas:	Parr
ISSN:	0022-3646
Additional Keywords:	CP12, GapC1, GAPDH, kinetics
NORA Subject Terms:	Botany Biology and Microbiology
Date made live:	01 Dec 2008 11:13 +0 (UTC)
URI:	https://nora.nerc.ac.uk/id/eprint/4436

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