OBJECTIVES: To develop a novel cerebrospinal fluid (CSF) beta-secretase-1 activity assay and evaluate beta-secretase-1 (BACE-1) activity as a potential biomarker in human Alzheimer's disease. METHODS: The assay consisted of an enzymatic reaction of CSF samples with an optimized beta-secretase peptide substrate and the cleavage products were detected using a neo-epitope specific antibody. RESULTS: The CSF BACE-1 activity assay described exhibits time, temperature, dose, and pH dependence, with sensitivity down to <1 pM of recombinant BACE-1 enzyme, and is completely blocked by BACE-1 inhibitors. The endogenous BACE-1 enzyme in CSF appears to exist as a c-terminally truncated protein, based on both western blotting and capture-based activity assays. In a small cohort of human subjects, an age-dependent increase in CSF BACE activity was observed (~1.0 pM/year, p<0.05). In Alzheimer's disease subjects, a significant decline in age-adjusted CSF BACE activity was observed compared to controls (56% in the log-transformed scale, p=0.02). CONCLUSION: We have developed a robust assay to measure CSF BACE-1 activity which could serve as a potential biomarker in human Alzheimer's disease subjects.
OBJECTIVES: To develop a novel cerebrospinal fluid (CSF) beta-secretase-1 activity assay and evaluate beta-secretase-1 (BACE-1) activity as a potential biomarker in humanAlzheimer's disease. METHODS: The assay consisted of an enzymatic reaction of CSF samples with an optimized beta-secretase peptide substrate and the cleavage products were detected using a neo-epitope specific antibody. RESULTS: The CSF BACE-1 activity assay described exhibits time, temperature, dose, and pH dependence, with sensitivity down to <1 pM of recombinant BACE-1 enzyme, and is completely blocked by BACE-1 inhibitors. The endogenous BACE-1 enzyme in CSF appears to exist as a c-terminally truncated protein, based on both western blotting and capture-based activity assays. In a small cohort of human subjects, an age-dependent increase in CSF BACE activity was observed (~1.0 pM/year, p<0.05). In Alzheimer's disease subjects, a significant decline in age-adjusted CSF BACE activity was observed compared to controls (56% in the log-transformed scale, p=0.02). CONCLUSION: We have developed a robust assay to measure CSF BACE-1 activity which could serve as a potential biomarker in humanAlzheimer's disease subjects.
Authors: L M Bekris; N M Galloway; S Millard; D Lockhart; G Li; D R Galasko; M R Farlow; C M Clark; J F Quinn; J A Kaye; G D Schellenberg; J B Leverenz; P Seubert; D W Tsuang; E R Peskind; C E Yu Journal: Neurobiol Aging Date: 2010-12-31 Impact factor: 4.673
Authors: Steven P Millard; Franziska Lutz; Ge Li; Douglas R Galasko; Martin R Farlow; Joseph F Quinn; Jeffrey A Kaye; James B Leverenz; Debby Tsuang; Chang-En Yu; Elaine R Peskind; Lynn M Bekris Journal: Neurobiol Aging Date: 2013-09-04 Impact factor: 4.673
Authors: Guoxin Wu; Sethu Sankaranarayanan; Donna L Montgomery; Adam J Simon; Zhiqiang An; Mary J Savage Journal: Protein Cell Date: 2011-08-06 Impact factor: 14.870
Authors: Inmaculada Lopez-Font; Claudia P Boix; Henrik Zetterberg; Kaj Blennow; Javier Sáez-Valero Journal: Mol Neurobiol Date: 2019-07-09 Impact factor: 5.590
Authors: Mary J Savage; Daniel J Holder; Guoxin Wu; June Kaplow; Judith A Siuciak; William Z Potter Journal: J Alzheimers Dis Date: 2015 Impact factor: 4.472