Universität zu Köln

Wulfert, Sabine, Schilasky, Soeren and Krueger, Stephan ORCID: 0000-0002-1658-5993 (2020). Transcriptional and Biochemical Characterization of Cytosolic Pyruvate Kinases in Arabidopsis thaliana. Plants-Basel, 9 (3). BASEL: MDPI. ISSN 2223-7747

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Abstract

Glycolysis is a central catabolic pathway in every living organism with an essential role in carbohydrate breakdown and ATP synthesis, thereby providing pyruvate to the tricarboxylic acid cycle (TCA cycle). The cytosolic pyruvate kinase (cPK) represents a key glycolytic enzyme by catalyzing phosphate transfer from phosphoenolpyruvate (PEP) to ADP for the synthesis of ATP. Besides its important functions in cellular energy homeostasis, the activity of cytosolic pyruvate kinase underlies tight regulation, for instance by allosteric effectors, that impact stability of its quaternary structure. We determined five cytosol-localized pyruvate kinases, out of the fourteen putative pyruvate kinase genes encoded by the Arabidopsis thaliana genome, by investigation of phylogeny and localization of yellow fluorescent protein (YFP) fusion proteins. Analysis of promoter beta-glucuronidase (GUS) reporter lines revealed an isoform-specific expression pattern for the five enzymes, subject to plant tissue and developmental stage. Investigation of the heterologously expressed and purified cytosolic pyruvate kinases revealed that these enzymes are differentially regulated by metabolites, such as citrate, fructose-1,6-bisphosphate (FBP) and ATP. In addition, measured in vitro enzyme activities suggest that pyruvate kinase subunit complexes consisting of cPK2/3 and cPK4/5 isoforms, respectively, bear regulatory properties. In summary, our study indicates that the five identified cytosolic pyruvate kinase isoforms adjust the carbohydrate flux through the glycolytic pathway in Arabidopsis thaliana, by distinct regulatory qualities, such as individual expression pattern as well as dissimilar responsiveness to allosteric effectors and enzyme subgroup association.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Wulfert, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED Schilasky, Soeren UNSPECIFIED UNSPECIFIED UNSPECIFIED Krueger, Stephan UNSPECIFIED orcid.org/0000-0002-1658-5993 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-342409
DOI:	10.3390/plants9030353
Journal or Publication Title:	Plants-Basel
Volume:	9
Number:	3
Date:	2020
Publisher:	MDPI
Place of Publication:	BASEL
ISSN:	2223-7747
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Botanical Institute
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language COLD-ACCLIMATION; GENE-EXPRESSION; PURIFICATION; GLYCOLYSIS; CASTOR; PHOTOSYNTHESIS; TRANSPORTERS; STRESS; MODEL; SEEDS Multiple languages Plant Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/34240