Myristoylation and its effects on the human Golgi Reassembly and Stacking Protein 55

Autor(es)

Kava, Emanuel ^[1] ; Mendes, Luis F. S. ^[1] ; Batista, Mariana R. B. ^{[1, 2]} ; Costa-Filho, Antonio J. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Ribeirao Preto Sch Philosophy Sci & Literature, Phys Dept, Mol Biophys Lab, Ribeirao Preto - Brazil

^[2] Univ Warwick, Sch Life Sci, Gibbet Hill Campus, Coventry CV4 7AL, W Midlands - England

• Tipo de documento: Artigo Científico
• Fonte: Biophysical Chemistry; v. 279, DEC 2021.
• Citações Web of Science: 0

Resumo

GRASP55 is a myristoylated protein localized in the medial/trans-Golgi faces and involved in the Golgi structure maintenance and the regulation of unconventional secretion pathways. It is believed that GRASP55 achieves its main functionalities in the Golgi organization by acting as a tethering factor. When bound to the lipid bilayer, its orientation relative to the membrane surface is restricted to determine its proper trans-oligomerization. Despite the paramount role of myristoylation in GRASP function, the impact of such protein modification on the membrane-anchoring properties and the structural organization of GRASP remains elusive. Here, an optimized protocol for the myristoylation in E. coli of the membrane-anchoring domain of GRASP55 is presented. The biophysical properties of the myristoylated/non-myristoylated GRASP55 GRASP domain were characterized in a membrane-mimicking micellar environment. Although myristoylation did not cause any impact on the protein's secondary structure, according to our circular dichroism data, it had a significant impact on the protein's thermal stability and solubility. Electrophoresis of negatively charged liposomes incubated with the two GRASP55 constructions showed different electrophoretic mobility for the myristoylated anchored protein only, thus demonstrating that myristoylation is essential for the biological membrane anchoring. Molecular dynamics simulations were used to further explore the anchoring process in determining the restricted orientation of GRASPs in the membrane. (AU)

• Processo FAPESP: 15/50366-7 - Resolving mechanistic details of peptide transport across membranes using crystallographic and non-crystallographic structural biology approaches
• Beneficiário: Antonio José da Costa Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/16328-3 - Simulações de dinâmica molecular de transportadores de oligopeptídeos dependentes de prótons
• Beneficiário: Mariana Raquel Bunoro Batista
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/24669-8 - Desvendando as bases moleculares de processos iniciais da secreção de proteínas em humanos utilizando técnicas biofísicas
• Beneficiário: Luis Felipe Santos Mendes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 12/20367-3 - Estudos estruturais e funcionais da proteína de organização e compactação do Golgi (GRASP) de Cryptococcus neoformans
• Beneficiário: Antonio José da Costa Filho
• Modalidade de apoio: Auxílio à Pesquisa - Regular