重组细菌L-天冬酰胺酶在人细胞中的表达。
Expression of a recombinant bacterial L-asparaginase in human cells.
作者信息
Dantas Raquel Caminha, Caetano Ludmilla Freire, Torres Ariany Lima Sousa, Alves Matheus Soares, Silva Emanuelly Thays Muniz Figueiredo, Teixeira Louhanna Pinheiro Rodrigues, Teixeira Daniel Câmara, de Azevedo Moreira Renato, Fonseca Marcela Helena Gambim, Gaudêncio Neto Saul, Martins Leonardo Tondello, Furtado Gilvan Pessoa, Tavares Kaio Cesar Simiano
机构信息
Experimental Biology Center (NUBEX), University of Fortaleza (UNIFOR), Fortaleza, Brazil.
Oswaldo Cruz Foundation (FIOCRUZ), Fortaleza, Brazil.
出版信息
BMC Res Notes. 2019 Dec 5;12(1):794. doi: 10.1186/s13104-019-4836-5.
OBJECTIVE
L-Asparaginase (ASNase) is an enzyme used in the treatment of acute lymphoblastic leukemia (ALL). As the therapeutic ASNases has bacterial origin, severe side effects are associated with its use, among them hypersensitivity and inactivation of the enzyme. In this context, the objective of this work was to produce a recombinant ASNase of bacterial origin in human cells in order to determine the presence and consequences of potential post-translational modifications on the enzyme.
RESULTS
Recombinant ASNase was expressed in human cells with a molecular weight of 60 kDa, larger than in Escherichia coli, which is 35 kDa. N-glycosylation analysis demonstrated that the increased molecular weight resulted from the addition of glycans to the protein by mammalian cells. The glycosylated ASNase presented in vitro activity at physiological pH and temperature. Given that glycosylation can act to reduce antigenicity by masking protein epitopes, our data may contribute to the development of an alternative ASNase in the treatment of ALL in patients who demonstrate side effects to currently marketed enzymes.
目的
L-天冬酰胺酶（ASNase）是一种用于治疗急性淋巴细胞白血病（ALL）的酶。由于治疗用的ASNase来源于细菌，使用该酶会产生严重的副作用，包括过敏反应和酶的失活。在此背景下，本研究的目的是在人细胞中生产一种细菌来源的重组ASNase，以确定潜在的翻译后修饰对该酶的影响及其后果。
结果
重组ASNase在人细胞中表达，分子量为60 kDa，大于在大肠杆菌中表达的35 kDa。N-糖基化分析表明，分子量的增加是由于哺乳动物细胞向该蛋白添加了聚糖。糖基化的ASNase在生理pH和温度下具有体外活性。鉴于糖基化可通过掩盖蛋白质表位来降低抗原性，我们的数据可能有助于开发一种替代ASNase，用于治疗对目前市场上的酶有副作用的ALL患者。
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