Brief introduction about journal related to functional amyloid called Pmel17 in mamalian cell and its function!

3. * Melanosome purification and extraction
* Sucrose buffer
+ 10mM Tris (pH 7.4)
+ 65mM NaCl
+ 2mM MgCl2
+ Protease Inhibitor cocktail
* Extraction buffer
+ 100mM Tris (pH 7.4)
+ 150mM NaCl
+ 0.1% NaN3

4. Fig1. B, C => purified melanosome all contained Pmel17
E, G => greater than 95% of melanosome fraction bound
thioflavin S and Congo Red
=> Melanosome contained Pmel17 amyloid.
Fig2. Melanin polymer is not responsible for fluorophore binding in
intact melanosomes, but Pmel17.
1μm 10μm
Insoluble clusters
Dense melanin-containing granule
lacking Pmel17
Pmel17-positive

5. α
* rMα (recombinant Mα) expression and purification

8. Full-length Pmel17 is synthesized and
trafficked to early melanosomes as a
transmembrane protein incapable of self-
assembly. Only when sequestered in the
specialized early melanosome compartment is
the amyloidogenic fragment, Mα, released by
proteolysis.
Importantly, the ability of Mα fibers to bind
and concentrate these reactive precursors
(DHQ) appears to protect the cell against the
toxicity caused by melanosome biogenesis.