Molecular cloning and NMR characterization of the nonreceptor tyrosine kinase PTK6 SH3-SH2-linker domain

Youngmin Lee, Kyo Eun Ahn, Sunggeon Ko, Weontae Lee

Research output: Contribution to journalArticle

Abstract

Human protein tyrosine kinase-6 (PTK6) is a member of the non-receptor protein tyrosine kinase family and it is found in two-thirds of all breast tumors. Very recently, we proposed that the SH3 domain of PTK6 interacts with the linker region (Linker) between the SH2 and kinase domains, proving that the interaction between SH3 domain and Linker plays an important role in auto-inhibition mechanism. Residues from 1 to 191 corresponding region of SH3-SH2-Linker (SH32L) of PTK6 was cloned into the pET32a expression vector with Tobbaco etch virus (TEV) protease enzyme site by sequence homology and 3D structural model. The purified PTK6-SH32L was determined as a monomer conformation in solution. The amide proton resonances in the 15N-1H 2D-HSQC spectrum suggest that PTK6-SH32L possesses disordered structural region of the flexible/unstructured linker region. In addition, the backbone amide proton chemical shifts of the SH3 domain in the PTK6-SH32L differ from that of the independent domain, indicating that intra-molecular interaction between SH3 and Linker in the PTK6-SH32L is present.

Original languageEnglish
Pages (from-to)1043-1046
Number of pages4
JournalBulletin of the Korean Chemical Society
Volume30
Issue number5
DOIs
Publication statusPublished - 2009 Oct 13

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Cloning
Protein-Tyrosine Kinases
Nuclear magnetic resonance
Amides
Protons
Molecular interactions
Chemical shift
Viruses
Conformations
Tumors
Peptide Hydrolases
Phosphotransferases
Monomers
Enzymes

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

Cite this

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title = "Molecular cloning and NMR characterization of the nonreceptor tyrosine kinase PTK6 SH3-SH2-linker domain",
abstract = "Human protein tyrosine kinase-6 (PTK6) is a member of the non-receptor protein tyrosine kinase family and it is found in two-thirds of all breast tumors. Very recently, we proposed that the SH3 domain of PTK6 interacts with the linker region (Linker) between the SH2 and kinase domains, proving that the interaction between SH3 domain and Linker plays an important role in auto-inhibition mechanism. Residues from 1 to 191 corresponding region of SH3-SH2-Linker (SH32L) of PTK6 was cloned into the pET32a expression vector with Tobbaco etch virus (TEV) protease enzyme site by sequence homology and 3D structural model. The purified PTK6-SH32L was determined as a monomer conformation in solution. The amide proton resonances in the 15N-1H 2D-HSQC spectrum suggest that PTK6-SH32L possesses disordered structural region of the flexible/unstructured linker region. In addition, the backbone amide proton chemical shifts of the SH3 domain in the PTK6-SH32L differ from that of the independent domain, indicating that intra-molecular interaction between SH3 and Linker in the PTK6-SH32L is present.",
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Molecular cloning and NMR characterization of the nonreceptor tyrosine kinase PTK6 SH3-SH2-linker domain. / Lee, Youngmin; Ahn, Kyo Eun; Ko, Sunggeon; Lee, Weontae.

In: Bulletin of the Korean Chemical Society, Vol. 30, No. 5, 13.10.2009, p. 1043-1046.

Research output: Contribution to journalArticle

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