Zinc binding regulates amyloid-like aggregation of GAPR-1
| Autor: | Sheng, Jie, Olrichs, Nick K, Geerts, Willie J, Li, Xueyi, Rehman, Ashfaq Ur, Gadella, Barend M, Kaloyanova, Dora V, Helms, J Bernd, LS Veterinaire biochemie, dB&C FR-RMSC FR, Sub Cryo - EM, dES/dFAH FR, Sub Reproductie mannelijk, Sub Biologie van de mannelijke gameet, dB&C FR-RMSC RMSC, Dep Biochemie en Celbiologie |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Conformational change Amyloid Biophysics Calorimetry Molecular Dynamics Simulation medicine.disease_cause Biochemistry CAP superfamily protein aggregation 03 medical and health sciences Mice Protein Domains Commentaries medicine Animals Humans Trypsin Salivary Proteins and Peptides Molecular Biology Pathogenesis-related protein membrane-interactions Mutation Binding Sites 030102 biochemistry & molecular biology Chemistry Heparin Circular Dichroism Seminal Plasma Proteins Membrane Proteins metal ions amyloid Isothermal titration calorimetry Cell Biology CRISP Protein tertiary structure GAPR-1 Zinc 030104 developmental biology Secretory protein glycosaminoglycans Commentary Sequence motif |
| Zdroj: | Bioscience Reports, 39(2). Portland Press Ltd. Bioscience Reports |
| ISSN: | 0144-8463 |
| Popis: | Members of the CAP superfamily (Cysteine-rich secretory proteins, Antigen 5, and Pathogenesis-related 1 proteins) are characterized by the presence of a CAP domain that is defined by four sequence motifs and a highly conserved tertiary structure. A common structure–function relationship for this domain is hitherto unknown. A characteristic of several CAP proteins is their formation of amyloid-like structures in the presence of lipids. Here we investigate the structural modulation of Golgi-Associated plant Pathogenesis Related protein 1 (GAPR-1) by known interactors of the CAP domain, preceding amyloid-like aggregation. Using isothermal titration calorimetry (ITC), we demonstrate that GAPR-1 binds zinc ions. Zn2+ binding causes a slight but significant conformational change as revealed by CD, tryptophan fluorescence, and trypsin digestion. The Zn2+-induced conformational change was required for the formation of GAPR-1 oligomers and amyloid-like assemblies in the presence of heparin, as shown by ThT fluorescence and TEM. Molecular dynamics simulations show binding of Zn2+ to His54 and His103. Mutation of these two highly conserved residues resulted in strongly diminished amyloid-like aggregation. Finally, we show that proteins from the cysteine-rich secretory protein (CRISP) subfamily are also able to form ThT-positive structures in vitro in a heparin- and Zn2+-dependent manner, suggesting that oligomerization regulated by metal ions could be a common structural property of the CAP domain. |
| Databáze: | OpenAIRE |
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