Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6-8

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Jun 1;63(Pt 6):480-3. doi: 10.1107/S1744309107020052. Epub 2007 May 5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Complement Factor H / biosynthesis
  • Complement Factor H / chemistry
  • Complement Factor H / genetics
  • Complement Factor H / isolation & purification
  • Crystallization
  • Crystallography, X-Ray
  • Gene Expression Regulation
  • Humans
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / isolation & purification
  • Sucrose / analogs & derivatives*
  • Sucrose / chemistry
  • Sucrose / isolation & purification
  • Sucrose / metabolism

Substances

  • CFH protein, human
  • Recombinant Proteins
  • Sucrose
  • Complement Factor H
  • sucrose octasulfate