The objective of this study was to determine the effect of growth rate on muscle fibre characteristics, concentration of nucleic acids (RNA and DNA) as indicators of muscle protein synthesis capacity and activity of the calpain system at time of slaughter in m. longissimus lumborum (LL) and m. supraspinatus (SS) from calves, in order to elucidate the effect of growth rate on muscle protein turn-over at time of slaughter. Twenty-four Friesian heifer calves were allocated to two different feeding regimens that allowed for a moderate/moderate (MM) or high/high (HH) growth rate from 5 days of age to 90 kg body weight (BW) (period I) and from 90 kg BW to slaughter at 250 kg BW (period II), respectively. The growth rates in the two periods and the weight of LL and SS at slaughter were recorded. Within 30 min after exsanguination, samples were removed from LL and SS, snap-frozen, and later analysed for muscle fibre type frequency and cross-sectional area, DNA and RNA concentration and the activity of the calpain system. High growth rate (i.e. 895 g/day and 1204 g/day in periods I and II, respectively), compared with moderate growth rate (678 g/day and 770 g/day in periods I and II, respectively) had a marked effect on muscle weight and muscle characteristics. High compared with moderate growth rate resulted in hypertrophic growth of type I, IIA and IIB fibres in LL and of type IIA and type IIB fibres in SS, but had no effect on the muscle fibre type frequency in either of the muscles. High growth rate increased total DNA and RNA content and the RNA: DNA ratio in LL, indicating a greater potential for protein synthesis in this muscle, whereas the effect of growth rate was smaller in SS. The activity of µ-calpain, m-calpain and calpastatin was higher in the red SS muscle compared with the whiter LL muscle. However, these enzyme activities were not affected by growth rate, and thus, did not indicate a higher myofibrillar proteolysis in vivo in calves exhibiting high growth rate compared with moderate growth rate. Overall the results showed that different types of muscles react differently to high versus moderate growth rate. High growth rate induced muscle hypertrophy and increased protein synthesis capacity especially in LL and less in SS, but the activities of the enzymes in the calpain system did not show any concomitant increase in muscle protein degradation that would be in favour of improved meat tenderness.