Biogenesis of gamma-secretase early in the secretory pathway.
Gamma-Secretase is responsible for proteolytic maturation of signaling and cell surface proteins, including amyloid precursor protein (APP). Abnormal processing of APP by gamma-secretase produces a fragment, Abeta(42), that may be responsible for Alzheimer's disease (AD). The biogenesis and trafficking of this important enzyme in relation to aberrant Abeta processing is not well defined. Using a cell-free reaction to monitor the exit of cargo proteins from the endoplasmic reticulum (ER), we have isolated a transient intermediate of gamma-secretase. Here, we provide direct evidence that the gamma-secretase complex is formed in an inactive complex at or before the assembly of an ER transport vesicle dependent on the COPII sorting subunit, Sec24A. Maturation of the holoenzyme is achieved in a subsequent compartment. Two familial AD (FAD)-linked PS1 variants are inefficiently packaged into transport vesicles generated from the ER. Our results suggest that aberrant trafficking of PS1 may contribute to disease pathology.
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA 94720, USA.
Jinoh
J
Kim
Bertrand
B
Kleizen
Regina
R
Choy
Gopal
G
Thinakaran
Sangram S
SS
Sisodia
Randy W
RW
Schekman
D053829:Amyloid Precursor Protein Secretases / Q000096:biosynthesis* / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D022181:COP-Coated Vesicles / Q000187:drug effects / Q000201:enzymology; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008744:Methylamines / Q000494:pharmacology; D050505:Mutant Proteins / Q000378:metabolism; D053764:Presenilin-1 / Q000096:biosynthesis; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D033921:Vesicular Transport Proteins / Q000378:metabolism
D008744:Methylamines; D050505:Mutant Proteins; D053764:Presenilin-1; C530638:SEC24A protein, human; D033921:Vesicular Transport Proteins; D053829:Amyloid Precursor Protein Secretases; C005855:trimethyloxamine