Molecular cloning and characterization of mouse cardiac triadin isoforms

Triadin is a ryanodine receptor and calsequestrin binding protein located in junctional sarcoplasmic reticulum of striated muscles. In the present study, mouse cardiac triadin cDNAs have been identified by cDNA library screening and RT-PCR. The deduced as sequences show that the three isoforms consist of 277, 293 and 305 as giving rise to the molecular weights of approximately 31,414, 33,066, and 34,328, respectively. The isoforms have identical 262 as N-terminal sequences, whereas they have distinct C-terminal sequences. Northern blot analysis using a cDNA probe representing the N-terminal common region of triadin revealed that the mouse triadins were present both in heart and skeletal muscles. The estimated sizes of the transcripts were approximately 1.3, 4.3 and 5 kb in heart and 5, 5.5 and 7 kb in skeletal muscle. Endo H treatment and Western blot analysis of isolated mouse cardiac sarcoplasmic reticulum and in vitro translation products indicate that there are three distinct mouse cardiac triadin isoforms having molecular weights of 35, 35.5 and 40 kDa. We termed those three isoforms as mouse cardiac triadin 1, mouse cardiac triadin 2 and mouse cardiac triadin 3. (C) 2001 Elsevier Science B.V. All rights reserved.