title: Structural studies of the transmembrane C-terminal domain of the amyloid precursor protein (APP): does APP function as a cholesterol sensor?
abstract: The amyloid precursor protein (APP) is subject to alternative pathways of proteolytic processing, leading either to production of the amyloid-beta (Abeta) peptides or to non-amyloidogenic fragments. Here, we report the first structural study of C99, the 99-residue transmembrane C-terminal domain of APP liberated by beta-secretase cleavage. We also show that cholesterol, an agent that promotes the amyloidogenic pathway, specifically binds to this protein. C99 was purified into model membranes where it was observed to homodimerize. NMR data show that the transmembrane domain of C99 is an alpha-helix that is flanked on both sides by mostly disordered extramembrane domains, with two exceptions. First, there is a short extracellular surface-associated helix located just after the site of alpha-secretase cleavage that helps to organize the connecting loop to the transmembrane domain, which is known to be essential for Abeta production. Second, there is a surface-associated helix located at the cytosolic C-terminus, adjacent to the YENPTY motif that plays critical roles in APP trafficking and protein-protein interactions. Cholesterol was seen to participate in saturable interactions with C99 that are centered at the critical loop connecting the extracellular helix to the transmembrane domain. Binding of cholesterol to C99 and, most likely, to APP may be critical for the trafficking of these proteins to cholesterol-rich membrane domains, which leads to cleavage by beta- and gamma-secretase and resulting amyloid-beta production. It is proposed that APP may serve as a cellular cholesterol sensor that is linked to mechanisms for suppressing cellular cholesterol uptake.
authors.affiliation: Department of Biochemistry and Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37232-8725, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Beel
authors.forename: Charles K
authors.initials: CK
authors.lastname: Mobley
authors.forename: Hak Jun
authors.initials: HJ
authors.lastname: Kim
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Arina
authors.initials: A
authors.lastname: Hadziselimovic
authors.forename: Bing
authors.initials: B
authors.lastname: Jap
authors.forename: James H
authors.initials: JH
authors.lastname: Prestegard
authors.forename: Charles R
authors.initials: CR
authors.lastname: Sanders
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology; D053829:Amyloid Precursor Protein Secretases / Q000737:chemistry / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism; D019281:Dimerization; D006801:Humans; D008567:Membranes, Artificial; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology
chemical_list: D016564:Amyloid beta-Protein Precursor; D008567:Membranes, Artificial; D010455:Peptides; D002784:Cholesterol; D053829:Amyloid Precursor Protein Secretases