Evidence map›Paper›PMID 37675505›Full record

ArticleAnnals of botany2023

Multiple highly expressed phosphoenolpyruvate carboxylase genes have divergent enzyme kinetic properties in two C4 grasses.

Robert J DiMario, Ashley N Kophs, Anthony J A Apalla, James N Schnable, Asaph B Cousins

Open access · bronzeAbstract read
In one paragraph

Article in Annals of botany, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
0.8field-weighted citation impact, top 27% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed, 5 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors at 2 institutions in 1 country.

Robert J DiMarioSchool of Biological Sciences, Washington State University, Pullman, WA 99164, USA.ORCID 0000-0002-5056-6868
Ashley N KophsSchool of Biological Sciences, Washington State University, Pullman, WA 99164, USA.
Anthony J A ApallaSchool of Biological Sciences, Washington State University, Pullman, WA 99164, USA.
James N SchnableDepartment of Agronomy and Horticulture, University of Nebraska, Lincoln, NE 68583, USA.
Asaph B CousinsSchool of Biological Sciences, Washington State University, Pullman, WA 99164, USA.ORCID 0000-0003-2424-714X
Washington State University · USUniversity of Nebraska–Lincoln · US

Funding

Department of Energy DE-SC0001685Division of Chemical Sciences, Geosciences, and BiosciencesNational Science Foundation 0923562Office of Basic Energy Sciences
6 · The paper itself

Abstract

BACKGROUND AND

aimsPhosphoenolpyruvate (PEP) carboxylase (PEPC) catalyses the irreversible carboxylation of PEP with bicarbonate to produce oxaloacetate. This reaction powers the carbon-concentrating mechanism (CCM) in plants that perform C4 photosynthesis. This CCM is generally driven by a single PEPC gene product that is highly expressed in the cytosol of mesophyll cells. We found two C4 grasses, Panicum miliaceum and Echinochloa colona, that each have two highly expressed PEPC genes. We characterized the kinetic properties of the two most abundant PEPCs in E. colona and P. miliaceum to better understand how the enzyme's amino acid structure influences its function.

methodsCoding sequences of the two most abundant PEPC proteins in E. colona and P. miliaceum were synthesized by GenScript and were inserted into bacteria expression plasmids. Point mutations resulting in substitutions at conserved amino acid residues (e.g. N-terminal serine and residue 890) were created via site-directed PCR mutagenesis. The kinetic properties of semi-purified plant PEPCs from Escherichia coli were analysed using membrane-inlet mass spectrometry and a spectrophotometric enzyme-coupled reaction. KEY

resultsThe two most abundant P. miliaceum PEPCs (PmPPC1 and PmPPC2) have similar sequence identities (>95 %), and as a result had similar kinetic properties. The two most abundant E. colona PEPCs (EcPPC1 and EcPPC2) had identities of ~78 % and had significantly different kinetic properties. The PmPPCs and EcPPCs had different responses to allosteric inhibitors and activators, and substitutions at the conserved N-terminal serine and residue 890 resulted in significantly altered responses to allosteric regulators.

conclusionsThe two, significantly expressed C4Ppc genes in P. miliaceum were probably the result of genomes combining from two closely related C4Panicum species. We found natural variation in PEPC's sensitivity to allosteric inhibition that seems to bypass the conserved 890 residue, suggesting alternative evolutionary pathways for increased malate tolerance and other kinetic properties.

Indexed as

Phosphoenolpyruvate CarboxylasePoaceaeAmino Acid SequenceBiological EvolutionKineticsPlantsSerinePhosphoenolpyruvate CarboxylaseSerineC4 photosynthesisEchinochloa colonaenzyme kineticsPanicum miliaceumphosphoenolpyruvate carboxylase

Identifiers

PMID37675505
PMCPMC10667006
OpenAlexW4386507585

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.