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The regulation of the thermal stability and affinity of the HSPA5 (Grp78/BiP) by clients and nucleotides is modulated by domains coupling.
Silva, Noeli S M; Siebeneichler, Bruna; Oliveira, Carlos S; Dores-Silva, Paulo R; Borges, Júlio C.
Afiliación
  • Silva NSM; São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP, Brazil. Electronic address: melo.ns@alumni.usp.br.
  • Siebeneichler B; São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP, Brazil; Exact and Technology Sciences Center, Federal University of São Carlos, São Carlos, SP 13560-970, Brazil.
  • Oliveira CS; São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP, Brazil.
  • Dores-Silva PR; São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP, Brazil.
  • Borges JC; São Carlos Institute of Chemistry, University of São Paulo, São Carlos, SP, Brazil. Electronic address: borgesjc@iqsc.usp.br.
Biochim Biophys Acta Proteins Proteom ; 1872(5): 141034, 2024 09 01.
Article en En | MEDLINE | ID: mdl-39009203
ABSTRACT
The HSPA5 protein (BiP/Grp78) serves as a pivotal chaperone in maintaining cellular protein quality control. As a member of the human HSP70 family, HSPA5 comprises two distinct domains a nucleotide-binding domain (NBD) and a peptide-binding domain (PBD). In this study, we investigated the interdomain interactions of HSPA5, aiming to elucidate how these domains regulate its function as a chaperone. Our findings revealed that HSPA5-FL, HSPA5-T, and HSPA5-N exhibit varying affinities for ATP and ADP, with a noticeable dependency on Mg2+ for optimal interactions. Interestingly, in ADP assays, the presence of the metal ion seems to enhance NBD binding only for HSPA5-FL and HSPA5-T. Moreover, while the truncation of the C-terminus does not significantly impact the thermal stability of HSPA5, experiments involving MgATP underscore its essential role in mediating interactions and nucleotide hydrolysis. Thermal stability assays further suggested that the NBD-PBD interface enhances the stability of the NBD, more pronounced for HSPA5 than for the orthologous HSPA1A, and prevents self-aggregation through interdomain coupling. Enzymatic analyses indicated that the presence of PBD enhances NBD ATPase activity and augments its nucleotide affinity. Notably, the intrinsic chaperone activity of the PBD is dependent on the presence of the NBD, potentially due to the propensity of the PBD for self-oligomerization. Collectively, our data highlight the pivotal role of allosteric mechanisms in modulating thermal stability, nucleotide interaction, and ATPase activity of HSPA5, underscoring its significance in protein quality control within cellular environments.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Adenosina Trifosfato / Estabilidad Proteica / Chaperón BiP del Retículo Endoplásmico / Proteínas de Choque Térmico Límite: Humans Idioma: En Revista: Biochim Biophys Acta Proteins Proteom Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
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Imprimir
XML
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Adenosina Trifosfato / Estabilidad Proteica / Chaperón BiP del Retículo Endoplásmico / Proteínas de Choque Térmico Límite: Humans Idioma: En Revista: Biochim Biophys Acta Proteins Proteom Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos