Journal of Protein Chemistry

, Volume 13, Issue 6, pp 517–525 | Cite as

Autophosphorylation-dependent protein kinase phosphorylates Ser25, Ser38, Ser65, Ser71, and Ser411 in vimentin and thereby inhibits cytoskeletal intermediate filament assembly

  • Tze-Jen Huang
  • Tsong-Tse Lee
  • Wen-Chuan Lee
  • Yiu-Kay Lai
  • Jau-Song Yu
  • Shiaw-Der Yang
Article
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Abstract

The autophosphorylation-dependent protein kinase has been identified as a potent vimentin kinase that incorporates 2 mol of phosphates per mol of protein and generates five major phosphorylation sites in vimentin. Tryptic phosphopeptide mapping by high-performance liquid chromatography followed by sequential manual Edman degradation and direct peptide sequence analysis revealed that Ser-25, Ser-38, Ser-65, and Ser-71 in the amino-terminal domain and Ser-411 in the carboxyl-terminal domain are the phosphorylation sites in vimentin phosphorylated by this kinase, indicating that autophosphorylation-dependent protein kinase is a potent and unique vimentin kinase. Functional study further revealed that phosphorylation of vimentin by autophosphorylation-dependent protein kinase can completely inhibit polymerization and assembly of the cytoskeletal intermediate filament as demonstrated by electron microscopic analysis. Taken together, the results provide initial evidence that the autophosphorylation-dependent protein kinase may function as a vimentin kinase involved in the structure-function regulation of the cytoskeletal system. The results also support the notion that this cyclic nucleotide- and calcium-independent protein kinase may function as a multisubstrate/multifunctional protein kinase involved in the regulation of diverse cell functions.

Key words

Autophosphorylation-dependent protein kinase vimentin assembly phosphorylation sites cytoskeletal intermediate filament 
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Copyright information

© Plenum Publishing Corporation 1994

Authors and Affiliations

  • Tze-Jen Huang
    • 1
    • 2
  • Tsong-Tse Lee
    • 1
    • 2
  • Wen-Chuan Lee
    • 1
    • 2
  • Yiu-Kay Lai
    • 1
    • 2
  • Jau-Song Yu
    • 1
    • 2
  • Shiaw-Der Yang
    • 1
    • 2
  1. 1.Institute of Biomedical and Life SciencesNational Tsing Hua UniversityHsinchuTaiwan, ROC
  2. 2.Institute of Basic MedicineChang Gung Medical CollegeTaiwan, ROC

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