Chloroplast outer envelope protein P39 in <scp><i>A</i></scp><i>rabidopsis thaliana</i> belongs to the Omp85 protein family

Hsueh, Yi‐Ching; Flinner, Nadine; Groß, Lucia E.; Haarmann, Raimund; Mirus, Oliver; Sommer, Maik S.; Schleiff, Enrico

doi:10.1002/prot.24725

Cited by 8 publications

(20 citation statements)

References 46 publications

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“…In brief, E. coli BL21 cells were transformed with the described plasmid (Hsueh et al . ). Expression of the protein was induced by addition of 1 mM IPTG at OD 600 = 1.…”

Section: Methodsmentioning

confidence: 97%

“…The purified full‐length His‐tagged protein was used for antibody generation as described previously (Hsueh et al . ).…”

Section: Methodsmentioning

confidence: 97%

“…The electrophysiological properties are comparable to those of Toc75‐V (Hsueh et al . ). At present, little is known about the function of p39, particularly in mature plants.…”

Section: Introductionmentioning

confidence: 97%

“…), namely the chloroplast‐localised At3 g44160 (p39; Hsueh et al . ) and At3 g48620 (p36; Nicolaisen et al . ).…”

Section: Introductionmentioning

confidence: 99%

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The outer membrane Omp85‐like protein P39 influences metabolic homeostasis in mature Arabidopsis thaliana

et al. 2018

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The Omp85 proteins form a large membrane protein family in bacteria and eukaryotes. Omp85 proteins are composed of a C-terminal β-barrel-shaped membrane domain and one or more N-terminal polypeptide transport-associated (POTRA) domains. However, Arabidopsis thaliana contains two genes coding for Omp85 proteins without a POTRA domain. One gene is designated P39, according to the molecular weight of the encoded protein. The protein is targeted to plastids and it was established that p39 has electrophysiological properties similar to other Omp85 family members, particularly to that designated as Toc75V/Oep80. We analysed expression of the gene and characterised two T-DNA insertion mutants, focusing on alterations in photosynthetic activity, plastid ultrastructure, global expression profile and metabolome. We observed pronounced expression of P39, especially in veins. Mutants of P39 show growth aberrations, reduced photosynthetic activity and changes in plastid ultrastructure, particularly in the leaf tip. Further, they display global alteration of gene expression and metabolite content in leaves of mature plants. We conclude that the function of the plastid-localised and vein-specific Omp85 family protein p39 is important, but not essential, for maintenance of metabolic homeostasis of full-grown A. thaliana plants. Further, the function of p39 in veins influences the functionality of other plant tissues. The link connecting p39 function with metabolic regulation in mature A. thaliana is discussed.

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“…In brief, E. coli BL21 cells were transformed with the described plasmid (Hsueh et al . ). Expression of the protein was induced by addition of 1 mM IPTG at OD 600 = 1.…”

Section: Methodsmentioning

confidence: 97%

“…The purified full‐length His‐tagged protein was used for antibody generation as described previously (Hsueh et al . ).…”

Section: Methodsmentioning

confidence: 97%

“…The electrophysiological properties are comparable to those of Toc75‐V (Hsueh et al . ). At present, little is known about the function of p39, particularly in mature plants.…”

Section: Introductionmentioning

confidence: 97%

“…), namely the chloroplast‐localised At3 g44160 (p39; Hsueh et al . ) and At3 g48620 (p36; Nicolaisen et al . ).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The outer membrane Omp85‐like protein P39 influences metabolic homeostasis in mature Arabidopsis thaliana

et al. 2018

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“…Similarly, the POTRA domain of Omp85 in the TOC complex, Toc75, recognizes the transit peptide of the chloroplast proteins that have to be translocated across the outer envelope and also interacts with another complex component, the receptor Toc34 (15). However, in Arabidopsis thaliana, two Omp85 homologs without the POTRA domain have been described experimentally (23,24), which is in line with the notion that Omp85 with either alternative or without soluble domains has evolved to perform alternative functions (25). POTRA domains might function as a hub for interaction with other complex partners, which might be absent in the case of the Omp85 proteins without POTRA domains.…”

Section: Introductionmentioning

confidence: 99%

Relative Orientation of POTRA Domains from Cyanobacterial Omp85 Studied by Pulsed EPR Spectroscopy

Dastvan

Brouwer

Schuetz

et al. 2016

Biophysical Journal

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Many proteins of the outer membrane of Gram-negative bacteria and of the outer envelope of the endosymbiotically derived organelles mitochondria and plastids have a b-barrel fold. Their insertion is assisted by membrane proteins of the Omp85-TpsB superfamily. These proteins are composed of a C-terminal b-barrel and a different number of N-terminal POTRA domains, three in the case of cyanobacterial Omp85. Based on structural studies of Omp85 proteins, including the five POTRAdomain-containing BamA protein of Escherichia coli, it is predicted that anaP2 and anaP3 bear a fixed orientation, whereas anaP1 and anaP2 are connected via a flexible hinge. We challenged this proposal by investigating the conformational space of the N-terminal POTRA domains of Omp85 from the cyanobacterium Anabaena sp. PCC 7120 using pulsed electron-electron double resonance (PELDOR, or DEER) spectroscopy. The pronounced dipolar oscillations observed for most of the double spinlabeled positions indicate a rather rigid orientation of the POTRA domains in frozen liquid solution. Based on the PELDOR distance data, structure refinement of the POTRA domains was performed taking two different approaches: 1) treating the individual POTRA domains as rigid bodies; and 2) using an all-atom refinement of the structure. Both refinement approaches yielded ensembles of model structures that are more restricted compared to the conformational ensemble obtained by molecular dynamics simulations, with only a slightly different orientation of N-terminal POTRA domains anaP1 and anaP2 compared with the x-ray structure. The results are discussed in the context of the native environment of the POTRA domains in the periplasm.

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Toc75‐V/OEP80 is processed during translocation into chloroplasts, and the membrane‐embedded form exposes its POTRA domain to the intermembrane space

et al. 2020

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The insertion of membrane proteins requires proteinaceous complexes in the cytoplasm, the membrane, and the lumen of organelles. Most of the required complexes have been described, while the components for insertion of β‐barrel‐type proteins into the outer membrane of chloroplasts remain unknown. The same holds true for the signals required for the insertion of β‐barrel‐type proteins. At present, only the processing of Toc75‐III, the β‐barrel‐type protein of the central chloroplast translocon with an atypical signal, has been explored in detail. However, it has been debated whether Toc75‐V/ outer envelope protein 80 (OEP80), a second protein of the same family, contains a signal and undergoes processing. To substantiate the hypothesis that Toc75‐V/OEP80 is processed as well, we reinvestigated the processing in a protoplast‐based assay as well as in native membranes. Our results confirm the existence of a cleavable segment. By protease protection and pegylation, we observed intermembrane space localization of the soluble N‐terminal domain. Thus, Toc75‐V contains a cleavable N‐terminal signal and exposes its polypeptide transport‐associated domains to the intermembrane space of plastids, where it likely interacts with its substrates.

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Chloroplast outer envelope protein P39 in Arabidopsis thaliana belongs to the Omp85 protein family

Cited by 8 publications

References 46 publications

The outer membrane Omp85‐like protein P39 influences metabolic homeostasis in mature Arabidopsis thaliana

The outer membrane Omp85‐like protein P39 influences metabolic homeostasis in mature Arabidopsis thaliana

Relative Orientation of POTRA Domains from Cyanobacterial Omp85 Studied by Pulsed EPR Spectroscopy

Toc75‐V/OEP80 is processed during translocation into chloroplasts, and the membrane‐embedded form exposes its POTRA domain to the intermembrane space

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