Document Detail


Effects of membrane lipids on the activity and processivity of purified γ-secretase.
MedLine Citation:
PMID:  22489600     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The 19-transmembrane multisubunit γ-secretase complex generates the amyloid β-peptide (Aβ) of Alzheimer's disease (AD) by intramembrane proteolysis of the β-amyloid precursor protein (APP). Despite substantial advances in elucidating how this protein complex functions, the effect of the local membrane lipid microenvironment on γ-secretase cleavage of substrates is still poorly understood. Using detergent-free proteoliposomes to reconstitute purified human γ-secretase, we examined the effects of fatty acyl (FA) chain length, saturation and double-bond isomerization, and membrane lipid polar headgroups on γ-secretase function. We analyzed γ-secretase activity and processivity [i.e., sequential cleavages in the APP transmembrane domain that convert longer Aβ species (e.g., Aβ(46)) into shorter ones (e.g., Aβ(40))] by quantifying the APP intracellular domain (AICD) and various Aβ peptides, including via a bicine/urea gel system that detects multiple Aβ lengths. These assays revealed several trends. (1) Switching from a cis to a trans isomer of a monounsaturated FA chain in phosphatidylcholine (PC) increased γ-activity, did not affect Aβ(42):Aβ(40) ratios, but decreased the ratio of long (≥42) versus short (≤41) Aβ peptides. (2) Increasing the FA carbon chain length (14, 16, 18, and 20) increased γ-activity, reduced longer Aβ species, and reduced the Aβ(42):Aβ(40) ratio. (3) Shifting the position of the double bond in 18:1(Δ9-cis) PC to the Δ6 position substantially reduced activity. (4) Gangliosides increased γ-activity but decreased processivity, thus elevating the Aβ(42):Aβ(40) ratio. (5) Phosphatidylserine decreased γ-activity but increased processivity. (6) Phosphatidylinositol strongly inhibited γ-activity. Overall, our results show that subtle changes in membrane lipid composition can greatly influence γ-secretase activity and processivity, suggesting that relatively small changes in lipid membrane composition may affect the risk of AD at least as much as presenilin or APP mutations do.
Authors:
Oliver Holmes; Swetha Paturi; Wenjuan Ye; Michael S Wolfe; Dennis J Selkoe
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-04-19
Journal Detail:
Title:  Biochemistry     Volume:  51     ISSN:  1520-4995     ISO Abbreviation:  Biochemistry     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-05-01     Completed Date:  2012-06-25     Revised Date:  2013-05-20    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3565-75     Citation Subset:  IM    
Affiliation:
Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, Massachusetts 02115, United States.
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MeSH Terms
Descriptor/Qualifier:
Amyloid Precursor Protein Secretases / isolation & purification*,  metabolism*
Amyloid beta-Peptides / chemistry,  metabolism
Animals
CHO Cells
Cricetinae
Enzyme Activation
Fatty Acids / chemistry
Humans
Lipid Bilayers / chemistry
Membrane Lipids / chemistry*,  metabolism
Peptide Fragments / chemistry,  metabolism
Protein Processing, Post-Translational*
Proteolysis
Substrate Specificity
Grant Support
ID/Acronym/Agency:
P01 AG015379/AG/NIA NIH HHS; P01 AG015379/AG/NIA NIH HHS; P01 AG015379-14/AG/NIA NIH HHS; R01 AG006173/AG/NIA NIH HHS; R01 AG012749/AG/NIA NIH HHS
Chemical
Reg. No./Substance:
0/Amyloid beta-Peptides; 0/Fatty Acids; 0/Lipid Bilayers; 0/Membrane Lipids; 0/Peptide Fragments; 0/amyloid beta-protein (1-40); 0/amyloid beta-protein (40-42); EC 3.4.-/Amyloid Precursor Protein Secretases
Comments/Corrections

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