Differential scanning calorimetry and CD spectrometry of acclimation temperature-associated types of carp light meromyosin.
Differential scanning calorimetry and CD spectrometry were employed to study the thermal unfolding of light meromyosin (LMM) prepared from carp acclimated to different temperatures. The transition temperatures given by the major peaks at pH 8.0 in 0.6 M KCl for LMM from carp acclimated to 10 degrees C were 32.5 and 39.5 degrees C with the calorimetric enthalpies (DeltaHcal) of 269 and 52 kcal/mol, respectively. LMM from carp acclimated to 20 degrees C exhibited three peaks of transition temperatures at 34.5, 40.2, and 46.9 with DeltaHcal of 152, 20, and 10 kcal/mol, respectively. On the other hand, LMM from carp acclimated to 30 degrees C showed two different patterns. The first experiment gave two transition temperatures at 39.2 and 47.3 degrees C with DeltaHcal of 231 and 39 kcal/mol, respectively. The second series of experiments resulted in showing three peaks of 34.4, 39.5, and 47.5 degrees C with DeltaHcal of 117, 123, and 28 kcal/mol, respectively. N-terminal amino acid sequence analysis revealed that LMM at the second series of experiments with the 30 degrees C-acclimated carp contained component(s) predominant in the 20 degrees C-acclimated carp. Thermal unfolding responsible for these transition temperatures was well explained by melting of alpha-helices which could be determined by far-ultraviolet CD spectroscopy. These results clearly demonstrate that the 30 degrees C-acclimated carp contained the most thermostable LMM.
Nakaya
M
M
Laboratory of Aquatic Molecular Biology and Biotechnology, University of Tokyo, Japan.
Kakinuma
M
M
Watabe
S
S
Ooi
T
T
D000064:Acclimatization; D000595:Amino Acid Sequence; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002347:Carps; D002942:Circular Dichroism; D008969:Molecular Sequence Data; D015879:Myosin Subfragments / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D013696:Temperature
D015879:Myosin Subfragments