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  • Yeast surface display of full-length human microtubule-associated protein tau.

Yeast surface display of full-length human microtubule-associated protein tau.

Biotechnology progress (2019-10-04)
Shiyao Wang, Yong Ku Cho
ABSTRACT

Microtubule-associated protein tau is an intrinsically disordered, highly soluble protein found primarily in neurons. Under normal conditions, tau regulates the stability of axonal microtubules and intracellular vesicle transport. However, in patients of neurodegeneration such as Alzheimer's disease (AD), tau forms neurofibrillary deposits, which correlates well with the disease progression. Identifying molecular signatures in tau, such as posttranslational modification, truncation, and conformational change has great potential to detect earliest signs of neurodegeneration and develop therapeutic strategies. Here, we show that full-length human tau, including the longest isoform found in the adult brain, can be robustly displayed on the surface of yeast Saccharomyces cerevisiae. Yeast-displayed tau binds to anti-tau antibodies that cover epitopes ranging from the N-terminus to the 4R repeat region. Unlike tau expressed in the yeast cytosol, surface-displayed tau was not phosphorylated at sites found in AD patients (probed by antibodies AT8, AT270, AT180, and PHF-1). However, yeast-displayed tau showed clear binding to paired helical filament (PHF) tau conformation-specific antibodies Alz-50, MC-1, and Tau-2. Although the tau possessed a conformation found in PHFs, oligomerization or aggregation into larger filaments was undetected. Taken together, yeast-displayed tau enables robust measurement of protein interactions and is of particular interest for characterizing conformational change.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Tau (4-repeat isoform RD4) Antibody, clone 1E1/A6, culture supernatant, clone 1E1/A6, Upstate®
Sigma-Aldrich
Anti-Tau Antibody, oligomeric Antibody, clone TOMA-1, clone TOMA-1, from mouse
Sigma-Aldrich
Anti-Tau-1 Antibody, clone PC1C6, clone PC1C6, Chemicon®, from mouse
Sigma-Aldrich
Anti-Tau antibody, Mouse monoclonal, clone Tau46, purified from hybridoma cell culture
Sigma-Aldrich
HRV-3C Protease., N-Terminal His tagged recombinant protein, aqueous solution, 0.8-1.2 mg/mL
Sigma-Aldrich
Monoclonal Anti-τ (Tau) antibody produced in mouse, clone TAU-2, ascites fluid
Hyperfilm ECL, W × L 5 in. × 7 in., pack of 50 ea