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Molecular and biochemical characterization of the parvulin-type PPIases in Lotus japonicus

Molecular and biochemical characterization of the parvulin-type PPIases in Lotus japonicus
Molecular and biochemical characterization of the parvulin-type PPIases in Lotus japonicus
The cis/trans isomerization of the peptide bond preceding proline is an intrinsically slow process, although important in many biological processes in both prokaryotes and eukaryotes. In vivo, this isomerization is catalyzed by peptidyl-prolyl cis/trans-isomerases (PPIases). Here, we present the molecular and biochemical characterization of parvulin-type PPIase family members of the model legume Lotus japonicus, annotated as LjPar1, LjPar2, and LjPar3. Although LjPar1 and LjPar2 were found to be homologous to PIN1 (Protein Interacting with NIMA)-type parvulins and hPar14 from human, respectively, LjPar3 represents a novel multidomain parvulin, apparently present only in plants, that contains an active carboxyl-terminal sulfurtransferase domain. All Lotus parvulins were heterologously expressed and purified from Escherichia coli, and purified protein verification measurements used a liquid chromatography-mass spectrometry-based proteomic method. The biochemical characterization of the recombinant Lotus parvulins revealed that they possess PPIase activity toward synthetic tetrapeptides, although they exhibited different substrate specificities depending on the amino acid amino terminal to proline. These differences were also studied in a structural context using molecular modeling of the encoded polypeptides. Real-time reverse transcription-polymerase chain reaction revealed that the three parvulin genes of Lotus are ubiquitously expressed in all plant organs. LjPar1 was found to be up-regulated during the later stages of nodule development. Subcellular localization of LjPar-enhanced Yellow Fluorescence Protein (eYFP) fusions expressed in Arabidopsis (Arabidopsis thaliana) leaf epidermal cells revealed that LjPar1- and LjPar2-eYFP fusions were localized in the cytoplasm and in the nucleus, in contrast to LjPar3-eYFP, which was clearly localized in plastids. Divergent substrate specificities, expression profiles, and subcellular localization indicate that plant parvulin-type PPIases are probably involved in a wide range of biochemical and physiological processes.
10.1104
0032-0889
1160-1173
Kouri, Evangelia D.
fcfa5414-a2d9-402b-b3b4-31523d54b342
Labrou, Nikolaos E.
6173f35e-27b3-44be-b158-1ad9e5cfd380
Garbis, Spiros D.
7067fd19-50c9-4d42-9611-f370289470bd
Kalliampakou, Katerina I.
1ba4baf7-3334-44a4-aeee-fd02e5c6c31d
Stedel, Catalina
fba9331c-a40e-4ba7-b798-d88cb6ca197c
Dimou, Maria
1191cdb3-8422-43d1-90d2-f1a6d85cb033
Udvardi, Michael K.
e36b798e-bfa6-47f0-9cc5-7cf7eb36f8b7
Katinakis, Panagiotis
cbee4488-9e98-422f-81c6-09a2fc2e2528
Flemetakis, Emmanouil
2bb962f6-53c4-4868-8096-8557ea030b14
Kouri, Evangelia D.
fcfa5414-a2d9-402b-b3b4-31523d54b342
Labrou, Nikolaos E.
6173f35e-27b3-44be-b158-1ad9e5cfd380
Garbis, Spiros D.
7067fd19-50c9-4d42-9611-f370289470bd
Kalliampakou, Katerina I.
1ba4baf7-3334-44a4-aeee-fd02e5c6c31d
Stedel, Catalina
fba9331c-a40e-4ba7-b798-d88cb6ca197c
Dimou, Maria
1191cdb3-8422-43d1-90d2-f1a6d85cb033
Udvardi, Michael K.
e36b798e-bfa6-47f0-9cc5-7cf7eb36f8b7
Katinakis, Panagiotis
cbee4488-9e98-422f-81c6-09a2fc2e2528
Flemetakis, Emmanouil
2bb962f6-53c4-4868-8096-8557ea030b14

Kouri, Evangelia D., Labrou, Nikolaos E., Garbis, Spiros D., Kalliampakou, Katerina I., Stedel, Catalina, Dimou, Maria, Udvardi, Michael K., Katinakis, Panagiotis and Flemetakis, Emmanouil (2009) Molecular and biochemical characterization of the parvulin-type PPIases in Lotus japonicus. Plant Physiology, 150 (3), 1160-1173. (doi:10.1104). (PMID:19403733)

Record type: Article

Abstract

The cis/trans isomerization of the peptide bond preceding proline is an intrinsically slow process, although important in many biological processes in both prokaryotes and eukaryotes. In vivo, this isomerization is catalyzed by peptidyl-prolyl cis/trans-isomerases (PPIases). Here, we present the molecular and biochemical characterization of parvulin-type PPIase family members of the model legume Lotus japonicus, annotated as LjPar1, LjPar2, and LjPar3. Although LjPar1 and LjPar2 were found to be homologous to PIN1 (Protein Interacting with NIMA)-type parvulins and hPar14 from human, respectively, LjPar3 represents a novel multidomain parvulin, apparently present only in plants, that contains an active carboxyl-terminal sulfurtransferase domain. All Lotus parvulins were heterologously expressed and purified from Escherichia coli, and purified protein verification measurements used a liquid chromatography-mass spectrometry-based proteomic method. The biochemical characterization of the recombinant Lotus parvulins revealed that they possess PPIase activity toward synthetic tetrapeptides, although they exhibited different substrate specificities depending on the amino acid amino terminal to proline. These differences were also studied in a structural context using molecular modeling of the encoded polypeptides. Real-time reverse transcription-polymerase chain reaction revealed that the three parvulin genes of Lotus are ubiquitously expressed in all plant organs. LjPar1 was found to be up-regulated during the later stages of nodule development. Subcellular localization of LjPar-enhanced Yellow Fluorescence Protein (eYFP) fusions expressed in Arabidopsis (Arabidopsis thaliana) leaf epidermal cells revealed that LjPar1- and LjPar2-eYFP fusions were localized in the cytoplasm and in the nucleus, in contrast to LjPar3-eYFP, which was clearly localized in plastids. Divergent substrate specificities, expression profiles, and subcellular localization indicate that plant parvulin-type PPIases are probably involved in a wide range of biochemical and physiological processes.

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e-pub ahead of print date: 29 April 2009
Published date: July 2009
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 395422
URI: http://eprints.soton.ac.uk/id/eprint/395422
ISSN: 0032-0889
PURE UUID: c2cd6537-b37e-43e6-a257-27f69bd217b6
ORCID for Spiros D. Garbis: ORCID iD orcid.org/0000-0002-1050-0805

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Date deposited: 18 Jul 2016 10:27
Last modified: 15 Mar 2024 00:39

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Contributors

Author: Evangelia D. Kouri
Author: Nikolaos E. Labrou
Author: Spiros D. Garbis ORCID iD
Author: Katerina I. Kalliampakou
Author: Catalina Stedel
Author: Maria Dimou
Author: Michael K. Udvardi
Author: Panagiotis Katinakis
Author: Emmanouil Flemetakis

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