Altered turnover of calcium regulatory proteins of the sarcoplasmic reticulum in aged skeletal muscle.
We have measured the in vivo protein turnover for the major calcium regulatory proteins of the sarcoplasmic reticulum from the skeletal muscle of young adult (7 months) and aged (28 months) Fischer 344 rats. From the time course of the incorporation and decay of protein-associated radioactivity after a pulse injection of [14C]leucine and correcting for leucine reutilization, in young rats, the apparent half-lives for calsequestrin, the 53-kDa glycoprotein, and ryanodine receptor are 5.4 +/- 0.4, 6.3 +/- 1.3, and 8.3 +/- 1.3 days, respectively. A half-life of 14.5 +/- 2.5 days was estimated for the Ca-ATPase isolated from young muscle. Differences in protein turnover associated with aging were determined using sequential injection of two different isotopic labels ([14C]leucine and [3H]leucine) to provide an estimate of protein synthesis and degradation within the same animal. The Ca-ATPase and ryanodine receptor isolated from aged muscle exhibits 27 +/- 5% and 25 +/- 3% slower protein turnover, respectively, relative to that from young muscle. In contrast, the 53-kDa glycoprotein exhibits a 25 +/- 5% more rapid turnover in aged SR, while calsequestrin exhibits no age-dependent alteration in turnover. Statistical analysis comparing the sensitivity of various methods for discriminating different rates of protein turnover validates the approach used in this study and demonstrates that the use of two isotopic labels provides at least a 6-fold more sensitive means to detect age-related differences in protein turnover relative to other methods.
Ferrington
D A
DA
Department of Biochemistry, Haworth Hall, University of Kansas, Lawrence, Kansas 66045, USA.
Krainev
A G
AG
Bigelow
D J
DJ
D000367:Age Factors; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002155:Calsequestrin / Q000032:analysis; D002250:Carbon Radioisotopes / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D007930:Leucine / Q000097:blood; D008297:Male; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis / Q000493:pharmacokinetics*; D018482:Muscle, Skeletal / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D014316:Tritium / Q000378:metabolism
D002155:Calsequestrin; D002250:Carbon Radioisotopes; D006023:Glycoproteins; D009124:Muscle Proteins; D019837:Ryanodine Receptor Calcium Release Channel; D014316:Tritium; D000252:Calcium-Transporting ATPases; D007930:Leucine; D002118:Calcium