title: Two complementary approaches for intracellular delivery of exogenous enzymes.
abstract: Intracellular delivery of biologically active proteins remains a formidable challenge in biomedical research. Here we show that biomedically relevant enzymes can be delivered into cells using a new DNA transfection reagent, lipofectamine 3000, allowing assessment of their intracellular functions. We also show that the J774.2 macrophage cell line exhibits unusual intracellular uptake of structurally and functionally distinct enzymes providing a convenient, reagent-free approach for evaluation of intracellular activities of enzymes.
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Aleksander
authors.initials: A
authors.lastname: Rust
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Hazirah H A
authors.initials: HH
authors.lastname: Hassan
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Sedelnikova
authors.affiliation: MRC laboratory of Molecular Biology, Hills Road, Cambridge, UK.
authors.forename: Dhevahi
authors.initials: D
authors.lastname: Niranjan
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Hautbergue
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Shaymaa A
authors.initials: SA
authors.lastname: Abbas
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Lynda
authors.initials: L
authors.lastname: Partridge
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: David
authors.initials: D
authors.lastname: Rice
authors.affiliation: Medizinische Hochschule Hannover, Carl-Neuberg-Straße, Hannover, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Binz
authors.affiliation: The University of Sheffield, Western Bank, Sheffield, UK.
authors.forename: Bazbek
authors.initials: B
authors.lastname: Davletov
mesh_terms: D000584:Amiloride / Q000494:pharmacology; D002452:Cell Count; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003911:Dextrans / Q000737:chemistry; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000494:pharmacology*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009928:Organ Specificity; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000494:pharmacology; D054789:Ribosome Inactivating Proteins, Type 1 / Q000737:chemistry / Q000494:pharmacology; D000076985:Saporins; D014162:Transfection / Q000379:methods*; D000097573:Red Fluorescent Protein
chemical_list: D000584:Amiloride; D003911:Dextrans; D016650:Fluorescein-5-isothiocyanate; D008081:Liposomes; D008164:Luminescent Proteins; D011500:Protein Synthesis Inhibitors; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D000097573:Red Fluorescent Protein; C015219:fluorescein isothiocyanate dextran