Expression, purification, and neurotrophic activity of amyloid precursor protein-secreted forms produced by yeast.
The secreted form of amyloid precursor protein (APPs) including most of the extracellular domain of APP is released from the cell surface, suggesting physiological significance of APPs in vivo. We used the methylotrophic yeast Pichia pastoris as a host system for the production of recombinant APPs (rAPPs). Two rAPPss derived from isoforms of APP (APP695 and APP770) were secreted into the culture medium from the yeast, which carried cDNA encoding the N-terminal portion of APP under the control of a P. pastoris alcohol oxidase promoter. Like APPss produced by the transfected COS-1 cells, the purified rAPPss from yeast were shown to be biologically active in terms of neurite outgrowth of embryonic rat neocortical explants. These rAPPss could be valuable tools for investigating the biological functions of APPss.
Ohsawa
I
I
Department of Neurochemistry, National Institute of Neuroscience, Tokyo, Japan.
Hirose
Y
Y
Ishiguro
M
M
Imai
Y
Y
Ishiura
S
S
Kohsaka
S
S
D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002540:Cerebral Cortex / Q000187:drug effects / Q000502:physiology*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004622:Embryo, Mammalian; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009838:Oligodeoxyribonucleotides; D009924:Organ Culture Techniques; D010843:Pichia; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D015183:Restriction Mapping; D014162:Transfection
D016564:Amyloid beta-Protein Precursor; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins