Physical characterization of serpin conformations.
The native serpin fold is characterized by being metastable. This thermodynamic characteristic is manifested in the conversion of the native state to other more stable conformations. Whilst this structural transition is required for proteinase inhibition and regulation of a range of biological phenomena, inappropriate structural changes can result in a number of disease states. Identification of these alternative conformations has been essential in our understanding of serpin structure and function. However, identifying these alternative forms is also important if we are not to misinterpret data due to the formation of these states during in vitro studies. The different physical properties of these alternative serpin conformational states make it possible to use a range of standard laboratory techniques to identify these structures. In this chapter, we will outline these general approaches that can be used routinely to identify the alternative serpin conformational states.
Dafforn
Timothy R
TR
School of Biological Sciences, University of Manchester, Manchester M13 9PT, UK.
Pike
Robert N
RN
Bottomley
Stephen P
SP
D001665:Binding Sites; D002850:Chromatography, Gel; D002942:Circular Dichroism / Q000379:methods; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D010450:Endopeptidases / Q000378:metabolism; D008958:Models, Molecular; D010455:Peptides / Q000378:metabolism; D011108:Polymers / Q000032:analysis / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D015843:Serpins / Q000737:chemistry* / Q000378:metabolism
D010455:Peptides; D011108:Polymers; D015843:Serpins; D010450:Endopeptidases