PEGylation versus glycosylation: effect on the thermodynamics and thermostability of crisantaspase

Autor(es)

Torres-Obreque, Karin ; Kleingesinds, Eduardo Krebs ; Santos, Joao H. P. M. ; Carretero, Gustavo ; Rabelo, Jheniffer ; Converti, Attilio ; Monteiro, Gisele ; Pessoa Jr, Adalberto ; Rangel-Yagui, Carlota O.

• Tipo de documento: Artigo Científico
• Fonte: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY; v. N/A, p. 11-pg., 2023-09-11.

Resumo

Thermostability is an important and desired feature of therapeutic proteins and is critical for the success or failure of protein drugs development. It can be increased by PEGylation-binding of poly(ethylene glycol) moieties-or glycosylation-post-translational modification to add glycans. Here, the thermostability and thermodynamic parameters of native, PEGylated, and glycosylated versions of the antileukemic enzyme crisantaspase were investigated. First-order kinetics was found to describe the irreversible deactivation process. Activation energy of the enzyme-catalyzed reaction (E*) was estimated for native, PEGylated, and glycosylated enzyme (10.2, 14.8, and 18.8 kJ mol(-1) respectively). Half-life decreased progressively with increasing temperature, and lon-ger half-life was observed for PEG-crisantaspase (87.74 min) at 50 degrees C compared to the native form (9.79 min). The activation energy of denaturation of PEG-crisantaspase (307.1 kJ mol(-1)) was higher than for crisantaspase (218.1 kJ mol(-1)) and Glyco-crisantaspase (120.0 kJ mol(-1)), which means that more energy is required to overcome the energy barrier of the unfolding process. According to our results, PEG-crisantaspase is more thermostable than its native form, while Glyco-crisantaspase is more thermosensitive. (AU)

• Processo FAPESP: 16/22065-5 - Pegilação N-terminal de proteínas e purificação por sistemas aquosos bifásicos
• Beneficiário: Carlota de Oliveira Rangel Yagui
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/20384-9 - Desenvolvimento de processo de purificação de L-asparaginase humanizada e sua caracterização
• Beneficiário: Eduardo Krebs Kleingesinds
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/08617-7 - Produção de L-asparaginase extracelular: da bioprospecção à engenharia de um biofármaco antileucêmico
• Beneficiário: Adalberto Pessoa Junior
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/25994-2 - Desenvolvimento de novas plataformas de PEGuilação de proteínas com potencial terapêutico com recurso à microfluídica
• Beneficiário: João Henrique Picado Madalena Santos
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado