Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6-8.
Human plasma protein complement factor H (FH) is an inhibitor of the spontaneously activated alternative complement pathway. An allotypic variant of FH, 402His, has been associated with age-related macular degeneration, the leading cause of blindness in the elderly. Crystals of FH domains 6-8 (FH678) containing 402His have been grown in the presence of a polyanionic sucrose octasulfate ligand (an analogue of the natural glycosaminoglycan ligands of FH) using both native and selenomethionine-derivatized protein. Native data sets diffracting to 2.3 A and SeMet data sets of up to 2.8 A resolution have been collected. An anomalous difference Patterson map reveals self- and cross-peaks from two incorporated Se atoms. The corresponding selenium substructure has been solved.
Prosser
Beverly E
BE
The Sir William Dunn School of Pathology, The University of Oxford, Oxford, England, UK.
Johnson
Steven
S
Roversi
Pietro
P
Clark
Simon J
SJ
Tarelli
Edward
E
Sim
Robert B
RB
Day
Antony J
AJ
Lea
Susan M
SM
D017242:Complement Factor H / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D005786:Gene Expression Regulation; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D013395:Sucrose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
C493495:CFH protein, human; D011994:Recombinant Proteins; D013395:Sucrose; D017242:Complement Factor H; C053081:sucrose octasulfate