Instrumentation for automated determination of protein stability.
A new instrument system has been developed that automatically carries out solvent and thermal denaturations of proteins using fluorescence as a probe of structure. This instrument also automatically performs pH titrations and can make kinetic measurements on the time scale of seconds. The design philosophy and implementation are described. The prototype instrument was subjected to extensive testing. The instrument can very reproducibly and accurately collect data that allow the calculation of the thermodynamics of protein denaturation. This data collection can proceed without human intervention after acquisition start until the data are saved.
Stites
W E
WE
Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville 72701-1201, USA.
Byrne
M P
MP
Aviv
J
J
Kaplan
M
M
Curtis
P M
PM
D019791:Guanidine; D006146:Guanidines / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008836:Micrococcal Nuclease / Q000737:chemistry; D011489:Protein Denaturation; D012815:Signal Processing, Computer-Assisted; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D013816:Thermodynamics; D017186:Titrimetry
D006146:Guanidines; D008836:Micrococcal Nuclease; D013381:Subtilisins; D019791:Guanidine