title: Differences in stability among the human apolipoprotein E isoforms determined by the amino-terminal domain.
abstract: Denaturation by guanidine-HCl, urea, or heating was performed on the common isoforms of human apolipoprotein (apo) E (apoE2, apoE3, and apoE4) and their 22-kDa and 10-kDa fragments in order to investigate the effects of the cysteine/arginine interchanges at residues 112 and 158. Previous physical characterization of apoE3 established that apoE contains two domains, the 10-kDa carboxyl-terminal and 22-kDa amino-terminal domains, which unfold independently and exhibit large differences in stability. However, the physical properties of apoE2, apoE3, and apoE4 have not been compared before. Analysis by circular dichroism showed that the different isoforms have identical alpha-helical contents and guanidine-HCl denaturation confirmed that the two domains unfold independently in all three isoforms. However, guanidine-HCl, urea, and thermal denaturation showed differences in stability among the 22-kDa amino-terminal fragments of the apoE isoforms (apoE4 < apoE3 < apoE2). Furthermore, guanidine-HCl denaturation monitored by circular dichroism and fluorescence suggested the presence of a folding intermediate in apoE, most prominently in apoE4. Thus, these studies reveal that the major isoforms of apoE, which are associated with different pathological consequences, exhibit significant differences in stability.
authors.affiliation: Gladstone Institute of Neurological Disease, Cardiovascular Research Institute, Department of Pathology, University of California, San Francisco, California 94110, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Morrow
authors.forename: M L
authors.initials: ML
authors.lastname: Segall
authors.forename: S
authors.initials: S
authors.lastname: Lund-Katz
authors.forename: M C
authors.initials: MC
authors.lastname: Phillips
authors.forename: M
authors.initials: M
authors.lastname: Knapp
authors.forename: B
authors.initials: B
authors.lastname: Rupp
authors.forename: K H
authors.initials: KH
authors.lastname: Weisgraber
mesh_terms: D001057:Apolipoproteins E / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D019791:Guanidine / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D006868:Hydrolysis; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D013917:Thrombin / Q000378:metabolism; D014508:Urea / Q000737:chemistry
chemical_list: D001057:Apolipoproteins E; D010446:Peptide Fragments; D020033:Protein Isoforms; D014508:Urea; D013917:Thrombin; D019791:Guanidine