PuSH - Publikationsserver des Helmholtz Zentrums München

Rahim, M.S.A.* ; Sommer, L.A.M.* ; Wacker, A.* ; Schaad, M.* ; Dames, S.A.

H-1, N-15, and C-13 chemical shift assignments of the micelle immersed FAT C-terminal (FATC) domains of the human protein kinases ataxia-telangiectasia mutated (ATM) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) fused to the B1 domain of streptococcal protein G (GB1).

Biomol. NMR Assign. 12, 149-154 (2018)
Postprint DOI PMC
Open Access Green
FAT C-terminal (FATC) is a circa 33 residue-long domain. It controls the kinase functionality in phosphatidylinositol-3 kinase-related kinases (PIKKs). Recent NMR- and CD-monitored interaction studies indicated that the FATC domains of all PIKKs can interact with membrane mimetics albeit with different preferences for membrane properties such as surface charge and curvature. Thus they may generally act as membrane anchoring unit. Here, we present the H-1, N-15, and C-13 chemical shift assignments of the DPC micelle immersed FATC domains of the human PIKKs ataxia-telangiectasia mutated (ATM, residues 3024-3056) and DNA protein kinase catalytic subunit (DNA-PKcs, residues 4096-4128), both fused to the 56 residue long B1 domain of Streptococcal protein G (GB1). Each fusion protein is 100 amino acids long and contains in the linking region between the GB1 tag and the FATC region a thrombin (LVPRGS) and an enterokinase (DDDDK) protease site. The assignments pave the route for the detailed structural characterization of the membrane mimetic bound states, which will help to better understand the role of the proper cellular localization at membranes for the function and regulation of PIKKs. The chemical shift assignment of the GB1 tag is useful for NMR spectroscopists developing new experiments or using GB1 otherwise for case studies in the field of in-cell NMR spectroscopy or protein folding. Moreover it is often used as purification tag. Earlier we showed already that GB1 does not interact with membrane mimetics and thus does not disturb the NMR monitoring of membrane mimetic interactions of attached proteins.
Altmetric
Weitere Metriken?
[➜Einloggen]
Zusatzinfos bearbeiten [➜Einloggen]
Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Ataxia Telangiectasia Mutated (atm) ; Dna-dependent Kinase Catalytic Subunit (dna-pkcs) ; Fatc ; Phosphatidylinositol-3 Kinase-related Kinases (pikks) ; B1 Domain Of Streptococcal Protein G (gb1) ; Chemical Shift Assignment; Triple-resonance Experiment; Phosphatidylinositol 3-kinase-related Kinases; Isotopically-enriched Proteins; Cellular-response; Membrane Anchor; Backbone Amide; Nmr; Rapamycin; Target; Spectroscopy
ISSN (print) / ISBN 1874-2718
e-ISSN 1874-270X
Zeitschrift Biomolecular NMR Assignments
Quellenangaben Band: 12, Heft: 1, Seiten: 149-154 Artikelnummer: , Supplement: ,
Verlag Springer
Verlagsort Dordrecht
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Structural Biology (STB)