caa a22 4500 475744519 CHVBK 20180406123509.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1023/A:1026479212350 doi (NATIONALLICENCE)springer-10.1023/A:1026479212350 Differences in the Denaturation Behavior of Ribonuclease A Induced by Temperature and Guanidine Hydrochloride [Elektronische Daten] [Ulrich Arnold, Renate Ulbrich-Hofmann] Moderate temperatures or low concentrations of denaturants diminish the catalytic activity of some enzymes before spectroscopic methods indicate protein unfolding. To discriminate between possible reasons for the inactivation of ribonuclease A, we investigated the influence of temperature and guanidine hydrochloride on its proteolytic susceptibility to proteinase K by determining the proteolytic rate constants and fragment patterns. The results were related to changes of activity and spectroscopic properties of ribonuclease A. With thermal denaturation, the changes in activity and in the rate constants of proteolytic degradation coincide and occur slightly before the spectroscopically observable transition. In the case of guanidine hydrochloride-induced denaturation, however, proteolytic resistance of ribonuclease A initially increases accompanied by a drastic activity decrease far before unfolding of the protein is detected by spectroscopy or proteolysis. In addition to ionic effects, a tightening of the protein structure at low guanidine hydrochloride concentrations is suggested to be responsible for ribonuclease A inactivation. Plenum Publishing Corporation, 2000 Ribonuclease A nationallicence limited proteolysis nationallicence temperature nationallicence guanidine hydrochloride nationallicence unfolding nationallicence Arnold Ulrich Department of Biochemistry/Biotechnology, Martin-Luther University Halle-Wittenberg, D-06120, Halle, Germany aut Ulbrich-Hofmann Renate Department of Biochemistry/Biotechnology, Martin-Luther University Halle-Wittenberg, D-06120, Halle, Germany aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/5(2000-07-01), 345-352 0277-8033 19:5<345 2000 19 10930 https://doi.org/10.1023/A:1026479212350 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1026479212350 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Arnold Ulrich Department of Biochemistry/Biotechnology, Martin-Luther University Halle-Wittenberg, D-06120, Halle, Germany aut NATIONALLICENCE 700 1- Ulbrich-Hofmann Renate Department of Biochemistry/Biotechnology, Martin-Luther University Halle-Wittenberg, D-06120, Halle, Germany aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/5(2000-07-01), 345-352 0277-8033 19:5<345 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer