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Direct Mass-Spectrometric Identification of Arg296 and Arg299 as the Methylation Sites of hnRNP K Protein for Methyltransferase PRMT1 - The Protein Journal

  • ️Lin, Chao-Hsiung
  • ️Wed Dec 27 2006

Protein methylation is one of the most important post-translational modifications that contribute to the diversity and complexity of proteome. Here we report the study of in vitro methylation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) with protein arginine methyltransferase 1 (PRMT1), as an enzyme, and S-adenosyl-l-methionine (SAM), as a methyl donor. The mass analysis of tryptic peptides of hnRNP K before and after methylation reveals the addition of four methyl groups in the residues 288–303. Tandem mass-spectrometric analysis of this peptide shows that both Arg296 and Arg299 are dimethylated. In addition, fragmentation analysis of such methylated arginines illustrate that they are both asymmetric dimethylarginines. Since Arg296 and Arg299 are located near the SH3-binding domains of hnRNP K, such methylation has the potential in regulating the interaction of hnRNP K with Src protein family. Our results provide crucial information for further functional study of hnRNP K methylation.

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Abbreviations

hnRNP K:

heterogeneous nuclear ribonucleoprotein K

PRMT1:

protein arginine methyltransferase 1

SAM:

S-adenosyl-l-methionine

MS:

mass-spectrometry

MALDI:

matrix-assisted laser desorption/ionization

TOF:

time-of-flight

GAR:

glycine/arginine-rich

MS/MS:

tandem mass-spectrometry

STAT1:

signal transducers and activators of transcription 1

Sam68:

Src-associated substrate during mitosis 68 kDa

KH:

K homology

KI:

K-protein-interactive

SH3:

Src homology 3

Trx:

thioredoxin

GST:

glutathione-S-transferase

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Acknowledgments

We thank Drs. Gu-Gang Chang and Ming-Ji Fann for critical reading of the manuscript and valuable suggestions. This work was supported in part by the National Science Council (NSC 94-120-M-010-002, 94-2627-M-007-002), Taiwan.

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Authors and Affiliations

  1. Institute of Biopharmaceutical Sciences, National Yang-Ming University, 155, Li-Nong Street, Sec.2, Taipei, 11221, Taiwan

    Yi-Ying Chiou & Wey-Jinq Lin

  2. Department of Education and Research, Taipei City Hospital, Taipei, 10381, Taiwan

    Wey-Jinq Lin, Shu-Ling Fu & Chao-Hsiung Lin

  3. Institute of Traditional Medicine, National Yang-Ming University, 155, Li-Nong Street, Sec.2, Taipei, 11221, Taiwan

    Shu-Ling Fu

  4. Faculty of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, 155, Li-Nong Street, Sec.2, Taipei, 11221, Taiwan

    Chao-Hsiung Lin

Authors

  1. Yi-Ying Chiou

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  2. Wey-Jinq Lin

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  3. Shu-Ling Fu

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  4. Chao-Hsiung Lin

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Correspondence to Chao-Hsiung Lin.

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Chiou, YY., Lin, WJ., Fu, SL. et al. Direct Mass-Spectrometric Identification of Arg296 and Arg299 as the Methylation Sites of hnRNP K Protein for Methyltransferase PRMT1. Protein J 26, 87–93 (2007). https://doi.org/10.1007/s10930-006-9049-9

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  • Published: 27 December 2006

  • Issue Date: February 2007

  • DOI: https://doi.org/10.1007/s10930-006-9049-9

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