skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution

Abstract

Human Protein tyrosine kinase 6 (PTK6) (EC:2.7.10.2), also known as the breast tumor kinase (BRK), is an intracellular non-receptor Src-related tyrosine kinase expressed in a majority of human breast tumors and breast cancer cell lines, but its expression is low or completely absent in normal mammary glands. In the recent past, several studies have suggested that PTK6 is a potential therapeutic target in cancer. To understand its structural and functional properties, the PTK6 kinase domain (PTK6-KD) gene was cloned, overexpressed in a baculo-insect cell system, purified and crystallized at room temperature. X-ray diffraction data to 2.33 Å resolution was collected on a single PTK6-KD crystal, which belonged to the triclinic space group P1. The Matthews coefficient calculation suggested the presence of four protein molecules per asymmetric unit, with a solvent content of ∼50%.The structure has been solved by molecular replacement and crystal structure data submitted to the protein data bank under the accession number 5D7V. This is the first report of apo PTK6-KD structure crystallized in DFG-in and αC-helix-out conformation. - Highlights: • We report the first crystal structure of the kinase domain of human PTK6. • >95% pure recombinant protein was purified in two-steps protocol: affinity chromatography followed by gelmore » filtration. • Kinase activity of the crystal construct is comparable to the full length enzyme. • This apo PTK6-KD is in DFG-in and αC-helix-out conformation.« less

Authors:
 [1]; ; ; ;  [2];  [1]
  1. Department of Biochemistry, University of Mysore, Mysore, 570005 (India)
  2. Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022 (India)
Publication Date:
OSTI Identifier:
22696580
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 478; Journal Issue: 2; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; COMMERCIAL BUILDINGS; CRYSTAL STRUCTURE; MAMMARY GLANDS; NEOPLASMS; PHOSPHOTRANSFERASES; RESOLUTION; SPACE GROUPS; TEMPERATURE RANGE 0273-0400 K; TYROSINE; X RADIATION; X-RAY DIFFRACTION

Citation Formats

Thakur, Manish Kumar, Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022, Kumar, Amit, Birudukota, Swarnakumari, Swaminathan, Srinivasan, Tyagi, Rajiv, Gosu, Ramachandraiah, and Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022. Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution. United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2016.07.121.
Thakur, Manish Kumar, Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022, Kumar, Amit, Birudukota, Swarnakumari, Swaminathan, Srinivasan, Tyagi, Rajiv, Gosu, Ramachandraiah, & Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022. Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution. United States. https://doi.org/10.1016/J.BBRC.2016.07.121
Thakur, Manish Kumar, Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022, Kumar, Amit, Birudukota, Swarnakumari, Swaminathan, Srinivasan, Tyagi, Rajiv, Gosu, Ramachandraiah, and Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022. Fri . "Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution". United States. https://doi.org/10.1016/J.BBRC.2016.07.121.
@article{osti_22696580,
title = {Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution},
author = {Thakur, Manish Kumar and Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022 and Kumar, Amit and Birudukota, Swarnakumari and Swaminathan, Srinivasan and Tyagi, Rajiv and Gosu, Ramachandraiah and Department of Structural Biology, Jubilant Biosys Ltd, Bangalore, 560022},
abstractNote = {Human Protein tyrosine kinase 6 (PTK6) (EC:2.7.10.2), also known as the breast tumor kinase (BRK), is an intracellular non-receptor Src-related tyrosine kinase expressed in a majority of human breast tumors and breast cancer cell lines, but its expression is low or completely absent in normal mammary glands. In the recent past, several studies have suggested that PTK6 is a potential therapeutic target in cancer. To understand its structural and functional properties, the PTK6 kinase domain (PTK6-KD) gene was cloned, overexpressed in a baculo-insect cell system, purified and crystallized at room temperature. X-ray diffraction data to 2.33 Å resolution was collected on a single PTK6-KD crystal, which belonged to the triclinic space group P1. The Matthews coefficient calculation suggested the presence of four protein molecules per asymmetric unit, with a solvent content of ∼50%.The structure has been solved by molecular replacement and crystal structure data submitted to the protein data bank under the accession number 5D7V. This is the first report of apo PTK6-KD structure crystallized in DFG-in and αC-helix-out conformation. - Highlights: • We report the first crystal structure of the kinase domain of human PTK6. • >95% pure recombinant protein was purified in two-steps protocol: affinity chromatography followed by gel filtration. • Kinase activity of the crystal construct is comparable to the full length enzyme. • This apo PTK6-KD is in DFG-in and αC-helix-out conformation.},
doi = {10.1016/J.BBRC.2016.07.121},
url = {https://www.osti.gov/biblio/22696580}, journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 2,
volume = 478,
place = {United States},
year = {2016},
month = {9}
}