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Protein-protein interactions of tandem affinity purified protein kinases from rice.

Rohila JS, Chen M, Chen S, Chen J, Cerny RL, Dardick C, Canlas P, Fujii H, Gribskov M, Kanrar S, Knoflicek L, Stevenson B, Xie M, Xu X, Zheng X, Zhu JK, Ronald P, Fromm ME - PLoS ONE (2009)

Bottom Line: Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score.In vivo phosphorylated sites were found for three of the PKs.A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.

View Article: PubMed Central - PubMed

Affiliation: Plant Science Initiative, University of Nebraska, Lincoln, NE, USA. jai.rohila@sdstate.edu

ABSTRACT
Eighty-eight rice (Oryza sativa) cDNAs encoding rice leaf expressed protein kinases (PKs) were fused to a Tandem Affinity Purification tag (TAP-tag) and expressed in transgenic rice plants. The TAP-tagged PKs and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by tandem mass spectrometry. Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score. In vivo phosphorylated sites were found for three of the PKs. A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.

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Sypro ruby stained TAP-purified protein complexes after SDS-PAGE.(A) TAP-purified rice Os01g14932 in-vivo protein complex from transgenic rice plants. (B) TAP-purified rice Os03g08550 in-vivo protein complex. The names of the proteins identified by mass spectrometry (MS) analysis of the digested peptides are shown in the bands. Standard protein molecular marker is shown on left side of the gel.
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pone-0006685-g002: Sypro ruby stained TAP-purified protein complexes after SDS-PAGE.(A) TAP-purified rice Os01g14932 in-vivo protein complex from transgenic rice plants. (B) TAP-purified rice Os03g08550 in-vivo protein complex. The names of the proteins identified by mass spectrometry (MS) analysis of the digested peptides are shown in the bands. Standard protein molecular marker is shown on left side of the gel.

Mentions: Additionally, PK1A-related Os05g02020 protein was found to interact with several members of the dynamin protein family [Dynamin-2A Os06g13820; dynamin-2B Os02g50550; dynamin-related protein 1A Os05g48240; dynamin-related protein 1C Os03g50520; and dynamin-related protein 1C Os10g41820 (Table 1, no. 7)]. Dynamin is a large GTPase protein involved in cell and organellar membrane budding, transport and fission [38]. This suggests that the PK1A-related Os05g02020 protein might be involved in regulating cellular membrane processes. Additional interacting proteins found include the RNA-related Ribonuclease T2 family protein Os09g36700 (Table 1, no. 7) (Figure 2B).


Protein-protein interactions of tandem affinity purified protein kinases from rice.

Rohila JS, Chen M, Chen S, Chen J, Cerny RL, Dardick C, Canlas P, Fujii H, Gribskov M, Kanrar S, Knoflicek L, Stevenson B, Xie M, Xu X, Zheng X, Zhu JK, Ronald P, Fromm ME - PLoS ONE (2009)

Sypro ruby stained TAP-purified protein complexes after SDS-PAGE.(A) TAP-purified rice Os01g14932 in-vivo protein complex from transgenic rice plants. (B) TAP-purified rice Os03g08550 in-vivo protein complex. The names of the proteins identified by mass spectrometry (MS) analysis of the digested peptides are shown in the bands. Standard protein molecular marker is shown on left side of the gel.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2723914&req=5

pone-0006685-g002: Sypro ruby stained TAP-purified protein complexes after SDS-PAGE.(A) TAP-purified rice Os01g14932 in-vivo protein complex from transgenic rice plants. (B) TAP-purified rice Os03g08550 in-vivo protein complex. The names of the proteins identified by mass spectrometry (MS) analysis of the digested peptides are shown in the bands. Standard protein molecular marker is shown on left side of the gel.
Mentions: Additionally, PK1A-related Os05g02020 protein was found to interact with several members of the dynamin protein family [Dynamin-2A Os06g13820; dynamin-2B Os02g50550; dynamin-related protein 1A Os05g48240; dynamin-related protein 1C Os03g50520; and dynamin-related protein 1C Os10g41820 (Table 1, no. 7)]. Dynamin is a large GTPase protein involved in cell and organellar membrane budding, transport and fission [38]. This suggests that the PK1A-related Os05g02020 protein might be involved in regulating cellular membrane processes. Additional interacting proteins found include the RNA-related Ribonuclease T2 family protein Os09g36700 (Table 1, no. 7) (Figure 2B).

Bottom Line: Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score.In vivo phosphorylated sites were found for three of the PKs.A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.

View Article: PubMed Central - PubMed

Affiliation: Plant Science Initiative, University of Nebraska, Lincoln, NE, USA. jai.rohila@sdstate.edu

ABSTRACT
Eighty-eight rice (Oryza sativa) cDNAs encoding rice leaf expressed protein kinases (PKs) were fused to a Tandem Affinity Purification tag (TAP-tag) and expressed in transgenic rice plants. The TAP-tagged PKs and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by tandem mass spectrometry. Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score. In vivo phosphorylated sites were found for three of the PKs. A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.

Show MeSH
Related in: MedlinePlus