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¹H, ¹⁵N and ¹³C backbone chemical shift assignment of titin domains A59-A60 and A60 alone

Lookup NU author(s): Professor Steve Homans

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Abstract

The giant protein titin is the third most abundant protein of vertebrate striated muscle. The titin molecule is > 1 mu m long and spans half the sarcomere, from the Z-disk to the M-line, and has important roles in sarcomere assembly, elasticity and intracellular signaling. In the A-band of the sarcomere titin is attached to the thick filaments and mainly consists immunoglobulin-like and fibronectin type III-like domains. These are mostly arranged in long-range patterns or 'super-repeats'. The large super-repeats each contain 11 domains and are repeated 11 times, thus forming nearly half the titin molecule. Through interactions with myosin and C-protein, they are involved in thick filament assembly. The importance of titin in muscle assembly is highlighted by the effect of mutations in the A-band portion, which are the commonest cause of dilated cardiomyopathy, affecting similar to 1 in 250 (Herman et al. in N Engl J Med 366:619-628, 2012). Here we report backbone N-15, C-13 and H-1 chemical shift and C-13 beta assignments for the A59-A60 domain tandem from the titin A59-A69 large super-repeat, completed using triple resonance NMR. Since, some regions of the backbone remained unassigned in A60 domain of the complete A59-A60 tandem, a construct containing a single A60 domain, A60sd, was also studied using the same methods. Considerably improved assignment coverage was achieved using A60sd due to its lower mass and improved molecular tumbling rate; these assignments also allowed the analysis of inter-domain interactions using chemical shift mapping against A59-A60.

Publication metadata

Author(s): Czajlik A, Thompson GS, Khan GN, Kalverda AP, Homans SW, Trinick J

Publication type: Article

Publication status: Published

Journal: Biomolecular NMR Assignments

Year: 2014

Volume: 8

Issue: 2

Pages: 429-433

Print publication date: 01/10/2014

Online publication date: 28/01/2014

Acceptance date: 12/12/2013

Date deposited: 29/10/2014

ISSN (print): 1874-2718

ISSN (electronic): 1874-270X

Publisher: Springer

URL: http://dx.doi.org/10.1007/s12104-013-9532-0

DOI: 10.1007/s12104-013-9532-0

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