Cited by 58Photosynthetic Processes and MechanismsBiochemical and Molecular ResearchEnzyme Structure and Function

Pyruvate kinase isozymes from the green alga, Selenastrum minutum

Lin, Min, Turpin, David H., Plaxton, William C.·1989·Archives of Biochemistry and Biophysics

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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Crossref · 10.1016/0003-9861(89)90103-3

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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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