Pyruvate kinase isozymes from the green alga, Selenastrum minutum
Abstract
Pyruvate kinase from the green alga Selenastrum minutum consists of two isoforms (PK1 and PK2) separable by Q-Sepharose chromatography. The two isoforms have been highly purified to respective final specific activities of 42 and 23 (mumol pyruvate produced/min)/mg protein. Purification steps included salt fractionation, anion-exchange, hydrophobic interaction, and gel filtration chromatography. The final enzyme preparations differ significantly in physical and immunological properties. PK1 is heat labile and is completely inactivated following reaction with N-ethylmaleimide. In contrast, PK2 is heat-stable and is only partially inactivated following N-ethylmaleimide treatment. PK1 appears to be homotetrameric with a native molecular mass of about 240 kDa, whereas PK2 appears to be homodecameric with a native molecular mass of approximately 590 kDa. The antigenic reaction of both final PK preparations to rabbit antiserum prepared against homogeneous germinating castor bean endosperm cytosolic pyruvate kinase was tested by immunoprecipitation and Western blotting. The two algal pyruvate kinases are immunologically unrelated as only PK2 cross-reacts with the cytosolic pyruvate kinase antibodies. These data indicate that the S. minutum pyruvate kinase isoforms, PK1 and PK2, are not interconvertible forms of the same protein, but probably represent chloroplastic and cytosolic isozymes, respectively.
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TCA intermediates usually suppress pyruvate kinase activity, the main modulating enzyme in carbohydrate catabolism.
“Usually, TCA intermediates cause the suppression of pyruvate kinase activity, the main modulating enzyme in carbohydrate catabolism (Lin et al. 1989).”
From Renewable energy from Cyanobacteria: energy production optimization by metabolic pathway engineering (2011) · cites this paper as Lin et al. 1989
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Renewable energy from Cyanobacteria: energy production optimization by metabolic pathway engineering
Quintana, N., Van der Kooy, F., Van de Rhee, M. D., Voshol, G. P., & Verpoorte, R. (2011) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-011-3394-0 · Cited by 338 · see citing works
- Authors
- Lin, M., Turpin, D. H., & Plaxton, W. C.
- Affiliations
- Queen's University (CA)
- Corresponding author
- Plaxton, William C.
- Venue
- Archives of Biochemistry and Biophysics
- Publisher
- Elsevier BV
- Published
- 1989-02
- Volume
- 269
- Issue
- 1
- Pages
- 219-227
- ISSN
- 0003-9861 (ISSN-L)
- OpenAlex
- W1580304949
- Cited by
- 58
- References
- 52
- Citations by year
- 2020: 1 · 2019: 1 · 2017: 2 · 2015: 1 · 2014: 1 · 2013: 1
- FWCI
- 1.42
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Archives of Biochemistry and Biophysics (publishedVersion)
- PubMed (publishedVersion)
- Funders
- West Virginia University
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Photosynthetic Processes and Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- pyruvate kinase, green algae, isozymes, enzyme purification, thermal stability, N-ethylmaleimide inactivation, quaternary structure, homotetramer, immunological cross-reactivity, algal metabolism
- MeSH terms
- Chlorophyta, Ammonium Sulfate, Animals, Chemistry, Physical, Chromatography, Agarose, Chromatography, High Pressure Liquid, Ethylmaleimide, Chemical Fractionation, Isoenzymes, Precipitin Tests, Pyruvate Kinase, Rabbits, Blotting, Western, Chemical Phenomena
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