title: Carboxylesterase 2 production and characterization in human cells: new insights into enzyme oligomerization and activity.
abstract: Carboxylesterase 2 (CES2), the main carboxylesterase expressed in human intestine, is an increasingly important enzyme in anti-cancer combined therapies for the treatment of different pathologies like colon adenocarcinoma and malignant glioma. The production of human recombinant CES2, in human embryonic kidney cells (HEK-293T cells) using serum-free media, is herein described. CES2 secretion to the media was achieved by the simple addition of an in-frame C-terminal 10× histidine tag (CES2-10xHis) without the need of addition of extra N-terminal signalling sequences or the mutation or deletion of the C-terminal HTEL motif responsible for retaining the protein in the lumen of endoplasmic reticulum. This secretion allowed a fourfold increase in CES2 production. The characterization of human recombinant CES2 showed that this protein exists in other active and inactive forms than the described 60 kDa monomer.
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157, Oeiras, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Lamego
authors.forename: Bárbara
authors.initials: B
authors.lastname: Cunha
authors.forename: Cristina
authors.initials: C
authors.lastname: Peixoto
authors.forename: Marcos F
authors.initials: MF
authors.lastname: Sousa
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.forename: Ana L
authors.initials: AL
authors.lastname: Simplício
authors.forename: Ana S
authors.initials: AS
authors.lastname: Coroadinha
mesh_terms: D043182:Carboxylesterase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D016895:Culture Media, Serum-Free / Q000737:chemistry; D006801:Humans; D008970:Molecular Weight; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D011993:Recombinant Fusion Proteins; C517204:CES2 protein, human; D043182:Carboxylesterase