Solution structure and backbone dynamics of the non-receptor protein-tyrosine kinase-6 Src homology 2 domain

Eunmi Hong, Joon Shin, Han Ie Kim, Seung-Taek Lee, Weon Tae Lee

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Human protein-tyrosine kinase-6 (PTK6, also known as breast tumor kinase (Brk)) is a member of the nonreceptor protein-tyrosine kinase family and is expressed in two-thirds of all breast tumors. To understand the structural basis of PTK6 function, we have determined the solution structure and backbone dynamics of the PTK6-Src homology 2 (SH2) domain using multidimensional NMR spectroscopy. The solution structure clearly indicates that the SH2 domain of human PTK6 contains a consensus α/β-fold and a Tyr(P) peptide binding surface, which are common to other SH2 domains. However, two of the α-helices (αA and αB) are located on opposite faces of the central β-sheet. In addition, the topological arrangement of a central four-stranded antiparallel β-sheet (strands βA, βB, γC, and βD) differs from that of other Src family members. Backbone dynamics and Tyr(P) peptide titration experiments revealed that the putative ligand binding sites of the PTK6-SH2 domain undergo distinctive internal motions when compared with other regions of the protein. Surface plasmon resonance analysis showed that the Tyr(P) peptide had a dissociation constant of about 60 μM, which is substantially weaker binding than previously reported for Src family members. The solution structure together with data from the ligand binding mode of PTK6-SH2 provides insight into the molecular basis of the autoinhibitory role of PTK6.

Original languageEnglish
Pages (from-to)29700-29708
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number28
DOIs
Publication statusPublished - 2004 Jul 9

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src Homology Domains
Protein-Tyrosine Kinases
Tumors
Ligands
Surface plasmon resonance
Breast Neoplasms
Titration
Nuclear magnetic resonance spectroscopy
Surface Plasmon Resonance
Phosphotransferases
Binding Sites
Consensus
Magnetic Resonance Spectroscopy
peptide P
Proteins
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

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title = "Solution structure and backbone dynamics of the non-receptor protein-tyrosine kinase-6 Src homology 2 domain",
abstract = "Human protein-tyrosine kinase-6 (PTK6, also known as breast tumor kinase (Brk)) is a member of the nonreceptor protein-tyrosine kinase family and is expressed in two-thirds of all breast tumors. To understand the structural basis of PTK6 function, we have determined the solution structure and backbone dynamics of the PTK6-Src homology 2 (SH2) domain using multidimensional NMR spectroscopy. The solution structure clearly indicates that the SH2 domain of human PTK6 contains a consensus α/β-fold and a Tyr(P) peptide binding surface, which are common to other SH2 domains. However, two of the α-helices (αA and αB) are located on opposite faces of the central β-sheet. In addition, the topological arrangement of a central four-stranded antiparallel β-sheet (strands βA, βB, γC, and βD) differs from that of other Src family members. Backbone dynamics and Tyr(P) peptide titration experiments revealed that the putative ligand binding sites of the PTK6-SH2 domain undergo distinctive internal motions when compared with other regions of the protein. Surface plasmon resonance analysis showed that the Tyr(P) peptide had a dissociation constant of about 60 μM, which is substantially weaker binding than previously reported for Src family members. The solution structure together with data from the ligand binding mode of PTK6-SH2 provides insight into the molecular basis of the autoinhibitory role of PTK6.",
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Solution structure and backbone dynamics of the non-receptor protein-tyrosine kinase-6 Src homology 2 domain. / Hong, Eunmi; Shin, Joon; Kim, Han Ie; Lee, Seung-Taek; Lee, Weon Tae.

In: Journal of Biological Chemistry, Vol. 279, No. 28, 09.07.2004, p. 29700-29708.

Research output: Contribution to journalArticle

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AU - Shin, Joon

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AU - Lee, Seung-Taek

AU - Lee, Weon Tae

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AB - Human protein-tyrosine kinase-6 (PTK6, also known as breast tumor kinase (Brk)) is a member of the nonreceptor protein-tyrosine kinase family and is expressed in two-thirds of all breast tumors. To understand the structural basis of PTK6 function, we have determined the solution structure and backbone dynamics of the PTK6-Src homology 2 (SH2) domain using multidimensional NMR spectroscopy. The solution structure clearly indicates that the SH2 domain of human PTK6 contains a consensus α/β-fold and a Tyr(P) peptide binding surface, which are common to other SH2 domains. However, two of the α-helices (αA and αB) are located on opposite faces of the central β-sheet. In addition, the topological arrangement of a central four-stranded antiparallel β-sheet (strands βA, βB, γC, and βD) differs from that of other Src family members. Backbone dynamics and Tyr(P) peptide titration experiments revealed that the putative ligand binding sites of the PTK6-SH2 domain undergo distinctive internal motions when compared with other regions of the protein. Surface plasmon resonance analysis showed that the Tyr(P) peptide had a dissociation constant of about 60 μM, which is substantially weaker binding than previously reported for Src family members. The solution structure together with data from the ligand binding mode of PTK6-SH2 provides insight into the molecular basis of the autoinhibitory role of PTK6.

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