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Acetylcholinesterase purification from human erythrocytes using magnetic nanoparticles containing procainamide.
Mororó, Maria Cleudenôra Cássia; Mahnke, Layla Carvalho; Assis, Caio Rodrigo Dias; da Silva, Roberto Afonso; Cabrera, Mariana Paola; Bezerra, Raquel Pedrosa; Carvalho Júnior, Luiz Bezerra; Alves, Maria Helena Menezes Estevam.
Afiliação
  • Mororó MCC; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil.
  • Mahnke LC; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil.
  • da Silva RA; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil.
  • Cabrera MP; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil; Departamento de Química Fundamental, Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, C
  • Bezerra RP; Departamento de Morfologia e Fisiologia Animal, Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil.
  • Carvalho Júnior LB; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil.
  • Alves MHME; Keizo Asami Intitute (iLIKA), Universidade Federal de Pernambuco, Avenida professor Moraes Rego, 1235, Cidade universitária, CEP 50670-901 Recife, Pernambuco, Brazil. Electronic address: maria@helena.bio.br.
Int J Biol Macromol ; 269(Pt 1): 132094, 2024 Jun.
Article em En | MEDLINE | ID: mdl-38705318
ABSTRACT
This work presents a magnetic purification method of human erythrocyte Acetylcholinesterase (EC 3.1.1.7; AChE) based on affinity binding to procainamide (Proca) as ligand. Acetylcholinesterase is an acetylcholine-regulating enzyme found in different areas of the body and associated with various neurological disorders, such as Parkinson, Alzheymer and Amyotrophic Lateral Sclerosis. AChE from human erythrocyte purification has been attempted in recent years with low degree of purity. Here, magnetic nanoparticles (MNP) were synthesized and coated with polyaniline (PANI) and procainamide (PROCA) was covalently linked to the PANI. The extracted human erythrocyte AChE formed a complex with the MNP@PANI-PROCA and an external magnet separated it from the undesired proteins. Finally, the enzyme was collected by increasing the ionic strength. Experimental Box-Behnken design was developed to optimize this process of human erythrocyte AChE purification protocol. The enzyme was purified in all fifteen experiments. However, the best AChE purification result was achieved, about 2000 times purified, when 100 mg of MNP@PANI-PROCA was incubated for one hour with 4 ml hemolysate extract. The SDS-PAGE of this preparation presented a molecular weight of approximately 70 kDa, corroborating with few previous studies of AChE from erythrocyte purification.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Procainamida / Acetilcolinesterase / Eritrócitos / Nanopartículas de Magnetita Limite: Humans Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Procainamida / Acetilcolinesterase / Eritrócitos / Nanopartículas de Magnetita Limite: Humans Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil