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29RNP kinase - Identification and characterization of chloroplast casein kinase II from Oryza sativa (rice)

ABSTRACT: We firstly identified a chloroplast casein kinase 2 from rice heparin sepharose enrichmen which could phosphorylate the RNP29. Then we identified the phosphorylation site of RNP29. We also performed the phosophoproteome analysis of rice and compared the phosphorylation site between rice and arabidopsis.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Oryza Sativa (rice)

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Qingtao Lu

LAB HEAD: Lu Qingtao

PROVIDER: PXD001168 | Pride | 2015-04-27

REPOSITORIES: Pride

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Identification and characterization of chloroplast casein kinase II from Oryza sativa (rice).

Lu Qingtao Q Ding Shunhua S Reiland Sonja S Rödiger Anja A Roschitzki Bernd B Xue Peng P Gruissem Wilhelm W Lu Congming C Baginsky Sacha S

Journal of experimental botany 20141014 1

Plastid casein kinase II is an important regulator of transcription, posttranscriptional processes, and, most likely, different metabolic functions in dicotyledonous species. Here we report the identification and characterization of pCKII from the monocotyledonous species Oryza sativa. OspCKII activity was enriched from isolated rice chloroplasts using heparin-Sepharose chromatography, in which it co-elutes with the transcriptionally active chromosome (TAC) and several ribosomal proteins. Inclus ...[more]

PMID: 25316064

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29RNP kinase idetification - Identification and characterization of chloroplast casein kinase II from Oryza sativa (rice)

Project description:The 29-kDa RNA protein (29RNP), which was originally identified in rice etioplasts, has increased phosphorylation levels during de-etiolation. Here, using heparin-sepharose enriched fractions, we identified a chloroplast kinase that phosphorylated 29RNP. The chloroplast kinase phosphorylated the 29RNP and exhibited CKII biochemical characteristics in a kinase reaction test. Proteomic analysis of the chloroplast heparin-sepharose enriched fractions revealed the presence of 70 proteins, including both abundant proteins, such as plastid encoded polymerase subunits, and low-abundance proteins, such as APO2 and DAG. An inclusion list method using Fourier transform ion cyclotron resonance mass spectrometer ( FT-ICR LTQ MS ) was subsequently used to analyze the chloroplast heparin-sepharose enriched fractions. The 29RNP kinase was positively identified as a CKII alpha family protein by the presence of two unique peptides.

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