Human apolipoprotein A-I: structure determination and analysis of unusual diffraction characteristics.
The crystallization and structure determination of recombinant human apolipoprotein A-I (apo A-I), the major protein component of high-density lipoprotein, is described. The fragment crystallized, residues 44-243 of native apo A-I [apo Delta(1-43)A-I], is very similar to intact native apo A-I in its ability to bind lipid, to be incorporated into high-density lipoproteins and to activate lecithin-cholesterol acyl transferase. Apo Delta(1-43)A-I crystallizes from 1.0-1.4 M sodium citrate pH 6.5-7.5 in space group P2(1)2(1)2(1), with unit-cell parameters a = 97.47, b = 113.87, c = 196.19 A (crystal form I). The crystals exhibit unusual diffraction intensity spikes and axial extinctions that are discussed in the context of the 4 A crystal structure. When flash-cooled to 100 K, the crystals diffract synchrotron radiation to 3 A resolution. Radiation sensitivity and crystal-to-crystal variation have hindered the assembly of a complete 3 A data set.
Borhani
D W
DW
Department of Organic Chemistry, Southern Research Institute, Birmingham, AL 35205, USA. borhani@sri.org
Engler
J A
JA
Brouillette
C G
CG
D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D055672:Static Electricity
D016632:Apolipoprotein A-I; D010446:Peptide Fragments; D011994:Recombinant Proteins