Photoincorporation of fluorescent probe into GroEL: defining site of interaction.
We have elucidated conditions for the covalent incorporation of a nonspecific hydrophobic probe, bisANS, into various proteins. Using this method, we are able to map hydrophobic surfaces in proteins. In addition, we have shown that for GroEL, we are able to use the fluorescence of the incorporated bisANS to monitor conformational changes in a defined region of the protein in response to various effectors. This method should be useful for studying both protein structure and dynamics.
Seale
J W
JW
Department of Biochemistry, University of Texas Health Science Center at San Antonio 78284-7760, USA.
Brazil
B T
BT
Horowitz
P M
PM
D000345:Affinity Labels; D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D001665:Binding Sites / Q000502:physiology; D018835:Chaperonin 10 / Q000378:metabolism; D018834:Chaperonin 60 / Q000737:chemistry*; D002918:Chymotrypsin / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D014466:Ultraviolet Rays
D000345:Affinity Labels; D000817:Anilino Naphthalenesulfonates; D018835:Chaperonin 10; D018834:Chaperonin 60; D005456:Fluorescent Dyes; D010446:Peptide Fragments; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D002918:Chymotrypsin