TY - JOUR
T1 - The contribution of individual residues of an aggregative hexapeptide derived from the human γD-crystallin to its amyloidogenicity
AU - Abu-Hussien, Malak
AU - Viswanathan, Guru Krishnakumar
AU - Simhaev, Luba
AU - Paul, Ashim
AU - Engel, Hamutal
AU - Gazit, Ehud
AU - Segal, Daniel
N1 - Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/3/15
Y1 - 2022/3/15
N2 - Human γD-crystallin protein is abundant in the lens and is essential for preserving lens transparency. With age the protein may lose its native structure resulting in the formation of cataract. We recently reported an aggregative peptide, 41Gly-Cys-Trp-Met-Leu-Tyr46 from the human γD-crystallin, termed GDC6, exhibiting amyloidogenic properties in vitro. Here, we aimed to determine the contribution of each residue of the GDC6 to its amyloidogenicity. Molecular dynamic (MD) simulations revealed that the residues Trp, Leu, and Tyr played an important role in the amyloidogenicity of GDC6 by facilitating inter-peptide main-chain hydrogen bonds, and π-π interactions. MD predictions were further validated using single-, double- and triple-alanine-substituted GDC6 peptides in which their amyloidogenic propensity was individually evaluated using complementary biophysical techniques including Thioflavin T assay, turbidity assay, CD spectroscopy, and TEM imaging. Results revealed that the substitution of Trp, Leu, and Tyr together by Ala completely abolished aggregation of GDC6 in vitro, highlighting their importance in the amyloidogenicity of GDC6.
AB - Human γD-crystallin protein is abundant in the lens and is essential for preserving lens transparency. With age the protein may lose its native structure resulting in the formation of cataract. We recently reported an aggregative peptide, 41Gly-Cys-Trp-Met-Leu-Tyr46 from the human γD-crystallin, termed GDC6, exhibiting amyloidogenic properties in vitro. Here, we aimed to determine the contribution of each residue of the GDC6 to its amyloidogenicity. Molecular dynamic (MD) simulations revealed that the residues Trp, Leu, and Tyr played an important role in the amyloidogenicity of GDC6 by facilitating inter-peptide main-chain hydrogen bonds, and π-π interactions. MD predictions were further validated using single-, double- and triple-alanine-substituted GDC6 peptides in which their amyloidogenic propensity was individually evaluated using complementary biophysical techniques including Thioflavin T assay, turbidity assay, CD spectroscopy, and TEM imaging. Results revealed that the substitution of Trp, Leu, and Tyr together by Ala completely abolished aggregation of GDC6 in vitro, highlighting their importance in the amyloidogenicity of GDC6.
KW - Amyloid aggregation
KW - Cataract
KW - GDC6 peptide
KW - Self-assembly
KW - γD-crystallin
UR - http://www.scopus.com/inward/record.url?scp=85122503766&partnerID=8YFLogxK
U2 - 10.1016/j.ijbiomac.2021.12.192
DO - 10.1016/j.ijbiomac.2021.12.192
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C2 - 34998884
AN - SCOPUS:85122503766
SN - 0141-8130
VL - 201
SP - 182
EP - 192
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
ER -