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Proteomic characteriaztion of the high light response of Calredoxin mutants (C. reinhardtii)

ABSTRACT: Calredoxin (CRX) is a calcium (Ca2+)-dependent thioredoxin (TRX) in the chloroplast of Chlamydomonas reinhardtii. To further elucidate the function of CRX, we performed in-depth quantitative proteomics taking advantage of a crx insertional mutant (IMcrx), two CRISPR/Cas9 KO mutants, CRX rescues and wild type strains. These analyses revealed that the chloroplast NADPH-dependent TRX reductase (NTRC) is co-regulated with CRX. Electron transfer measurements revealed that CRX inhibits NADPH-dependent reduction of oxidized chloroplast 2-Cys peroxiredoxin (PRX1) via NTRC and that the function of the NADPH-NTRC complex is under strict control of CRX. Our data also demonstrated via non-reducing SDS-PAGE assays and redox proteomics that PRX1, Transketolase (TRK1), ATPC and Proton Gradient Related like 1 (PGRL1) are more oxidized under high light (HL) conditions in the absence of CRX. The redox tuning of PRX1 and control of the NADPH-NTRC complex via CRX interconnects redox control with active photosynthetic electron transport and metabolism as well as Ca2+ signaling. In this way, an economic use of NADPH for PRX1 reduction is assured. The finding, that in the absence of CRX, light-driven CO2 fixation is severely inhibited under HL conditions underpins the importance of CRX for redox tuning as well as for efficient photosynthesis.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Chlamydomonas Reinhardtii

SUBMITTER: Martin Scholz

LAB HEAD: Michael Hippler

PROVIDER: PXD038009 | Pride | 2023-10-04

REPOSITORIES: Pride

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			Action	DRS
	2021-03-31-decoys-Creinhardtii_281_v5.6_Mito_Chloro_cRAP_29052020.fasta	Fasta
	GMM_20210510_HL-A5-1.mzML	Mzml
	GMM_20210510_HL-A5-1.raw	Raw
	GMM_20210510_HL-A5-2.mzML	Mzml
	GMM_20210510_HL-A5-2.raw	Raw

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