Pathogenic alpha 1-antitrypsin polymers are formed by reactive loop-beta-sheet A linkage.
alpha(1)-Antitrypsin is the most abundant circulating protease inhibitor and the archetype of the serine protease inhibitor or serpin superfamily. Members of this family may be inactivated by point mutations that favor transition to a polymeric conformation. This polymeric conformation underlies diseases as diverse as alpha(1)-antitrypsin deficiency-related cirrhosis, thrombosis, angio-edema, and dementia. The precise structural linkage within a polymer has been the subject of much debate with evidence for reactive loop insertion into beta-sheet A or C or as strand 7A. We have used site directed cysteine mutants and fluorescence resonance energy transfer (FRET) to measure a number of distances between monomeric units in polymeric alpha(1)-antitrypsin. We have then used a combinatorial approach to compare distances determined from FRET with distances obtained from 2.9 x 10(6) different possible orientations of the alpha(1)-antitrypsin polymer. The closest matches between experimental FRET measurements and theoretical structures show conclusively that polymers of alpha(1)-antitrypsin form by insertion of the reactive loop into beta-sheet A.
Respiratory Medicine Unit, Department of Medicine, University of Cambridge, The Wellcome Trust Centre for Molecular Mechanisms in Disease, Cambridge Institute for Medical Research, Cambridge, CB2 2XY, United Kingdom.
P
P
Sivasothy
T R
TR
Dafforn
P G
PG
Gettins
D A
DA
Lomas
D005453:Fluorescence; D008958:Models, Molecular; D011108:Polymers / Q000737:chemistry; D017433:Protein Structure, Secondary; D000515:alpha 1-Antitrypsin / Q000737:chemistry*
D011108:Polymers; D000515:alpha 1-Antitrypsin