TY - JOUR
T1 - Improving the heat resistance of ribonuclease A by the addition of poly(N,N-diethylaminoethyl methacrylate)-graft-poly(ethylene glycol) (PEAMA-g-PEG)
AU - Ganguli, Sumon
AU - Yoshimoto, Keitaro
AU - Tomita, Shunsuke
AU - Sakuma, Hiroshi
AU - Matsuoka, Tsuneyoshi
AU - Shiraki, Kentaro
AU - Nagasaki, Yukio
PY - 2010/8/11
Y1 - 2010/8/11
N2 - Poly(N,N-diethylaminoethyl methacrylate)-graft-poly(ethylene glycol) (PEAMA-g-PEG) has previously been used as a novel additive to improve the heat resistance of lysozyme, which has a positive net charge and a negatively charged active site. In the present study, we show that PEAMA-g-PEG prevents heat inactivation of ribonuclease A (RNase A), which has a positive net charge and a positively charged active site. After treatment at 98 °C for 10 min, the enzymatic activity of RNase A complexed with PEAMA-g-PEG was maintained at up to 75% of the level of the native RNase A. The extents of inactivation of RNase A and the complex of RNase A with PEAMA-g-PEG were strongly dependent upon the heating temperature and incubation time. Circular dichroism (CD) spectral analysis revealed that heat-induced irreversible inactivation was largely suppressed when RNase A was complexed with PEAMA-g-PEG. These findings suggest that the heat resistance of RNase A is improved by the external addition of PEAMA-g-PEG.
AB - Poly(N,N-diethylaminoethyl methacrylate)-graft-poly(ethylene glycol) (PEAMA-g-PEG) has previously been used as a novel additive to improve the heat resistance of lysozyme, which has a positive net charge and a negatively charged active site. In the present study, we show that PEAMA-g-PEG prevents heat inactivation of ribonuclease A (RNase A), which has a positive net charge and a positively charged active site. After treatment at 98 °C for 10 min, the enzymatic activity of RNase A complexed with PEAMA-g-PEG was maintained at up to 75% of the level of the native RNase A. The extents of inactivation of RNase A and the complex of RNase A with PEAMA-g-PEG were strongly dependent upon the heating temperature and incubation time. Circular dichroism (CD) spectral analysis revealed that heat-induced irreversible inactivation was largely suppressed when RNase A was complexed with PEAMA-g-PEG. These findings suggest that the heat resistance of RNase A is improved by the external addition of PEAMA-g-PEG.
UR - https://www.scopus.com/pages/publications/77956541777
UR - https://www.scopus.com/pages/publications/77956541777#tab=citedBy
U2 - 10.1002/mabi.200900432
DO - 10.1002/mabi.200900432
M3 - Article
C2 - 20301119
AN - SCOPUS:77956541777
SN - 1616-5187
VL - 10
SP - 853
EP - 859
JO - Macromolecular Bioscience
JF - Macromolecular Bioscience
IS - 8
ER -