Use of fluorescence resonance energy transfer to study serpin-proteinase interactions.
One of the important questions in the serpin mechanism of inhibition of serine and cysteine proteinases of different specificities and structural classes is whether a common "crushing" mechanism of proteinase inactivation is used in all cases. This mechanism was seen in an X-ray structure of the complex between alpha(1)-proteinase inhibitor and trypsin and required the full insertion of the reactive center loop into beta-sheet A and translocation of the proteinase from one pole of the serpin to the other. However, it has yet to be shown to be general for serine proteinases of structural classes other than the trypsin-fold or for cysteine proteinases with the papain-fold or for the caspases. Fluorescence resonance energy transfer offers a potential means of obtaining an answer to this question for each of these classes, without the concern for the effect that increasing size has on the observed signal that applies to NMR spectroscopy. However, care must be taken to ensure that measurements made represent sufficient overdetermination that the answer obtained is unambiguous.
Gettins
Peter G W
PG
Department of Biochemistry and Molecular Genetics, M/C 669, University of Illinois at Chicago, Chicago, IL 60607, USA. pgettins@uic.edu
Olson
Steven T
ST
D001665:Binding Sites; D018360:Crystallography, X-Ray; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D015843:Serpins / Q000737:chemistry / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000378:metabolism
D005456:Fluorescent Dyes; D015843:Serpins; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; C038915:alpha 1-antitrypsin Pittsburgh; D010450:Endopeptidases; D014357:Trypsin