By X. Wehrens, A. Marks
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Extra info for Arrhythmias and sudden death in athletes
Proteolytic digestion of RyRl yielded several major fragments,80-82 identified by sequencing80,82 or by immunoblotting with a series of seven antibodies81. Five major fragments were estimated to span the molecule. They had masses and deduced aa sequence boundaries as follows: 135 kDa (aa 1-1397 or 1508), 100 kDa (1398 or 1509-2401), 50 kDa (24022840), 160 kDa (3119-4475) and 76 kDa (4476-5037). 81 The more C-terminal 160 kDa fragment reacted with an antibody against an epitope contained within aa 4382-4417 and was resistant to extraction,81 implying that TM sequences exist in the sequence between aa 3119 and 4476.
The masses and approximate aa sequence of these fragments, beginning at the N-terminus are: 40 kDa (aa 1-426); 110 kDa (aa 426-1508); 100 kDa (aa 1508-2401); 50 kDa (aa 2401-2840); 33 kDa (aa 2840-3119); 150 kDa (aa 3119-4475); and 76 kDa (4475-5037). According to the hydropathy analysis and evidence obtained from other studies, the N- and C-termini of RyR1 are both located in the cytoplasm,83-85 indicating that an even number of sequences traverse the membrane. Antibodies against synthetic peptides corresponding to N-terminal aa 2-1584 and C-terminal aa 5027-503784 or 4941-503783 in rabbit skeletal muscle RyR1 bound to intact SR, confirming that both ends of the membraneembedded RyR1 are exposed to the cytoplasm.
ACKNOWLEDGEMENTS Original work from our laboratory, described in this review, was supported by grant MT-3399 to D. H. M. from the Canadian Institutes of Health Research. Chapter 3 THREE-DIMENSIONAL RECONSTRUCTION OF RYANODINE RECEPTORS Zheng Liu1 and Terence Wagenknecht1,2 1 Wadsworth Center, New York State Dept. of Health, Albany, New York; 2Dept. 96,97 One of the major calcium channels responsible for release from intracellular stores is the ryanodine receptor (RyR). RyRs are highly enriched in skeletal and cardiac muscle, or more precisely, in those regions of the sarcoplasmic reticulum (SR) membrane that interact with the plasma membrane/transverse tubule system (see Chapter 4).
Arrhythmias and sudden death in athletes by X. Wehrens, A. Marks