Expression of human extracellular superoxide dismutase in Chinese hamster ovary cells and characterization of the product

Journal Article · Thu Oct 01 04:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.19.6634· OSTI ID:7143888

Tibell, L; Hjalmarsson, K; Edlund, T; Skogman, G; Engstroem, A; Marklund, S L

A complementary DNA clone from human placenta, encoding human extracellular superoxide dismutase, has recently been isolated and characterized. An expression plasmid, based on the EC-SOD complementary DNA, was transfected into Chinese hamster ovary cells (CHO-K1). The transfected cells secreted human EC-SOD to the culture medium. The secreted recombinant (r) EC-SOD was isolated in high yield with a three-step procedure beginning with immobilized monoclonal anti-EC-SOD antibodies. The properties of the rEC-SOD were compared with native (n) EC-SOD isolated from human umbilical cords. The specific activities and amino-terminal amino acid sequences were identical. The amino acid compositions were virtually identical and very similar to the composition deduced from the complementary DNA sequence. Both rEC-SOD and nEC-SOD contained 4 Cu and 4 Zn atoms per molecule, and the presence of Zn in EC-SOD is thus now established. The rEC-SOD produced is type C, since its affinity for heparin-Sepharose was identical to that of nEC-SOD type C. Both enzymes bound to concanavalin A, lentil lectin, and wheat germ lectin and are thus glycoproteins. rEC-SOD and nEC-SOD seem to have the same subunit structure and composition as analyzed by polyacrylamide gel electrophoresis and gel chromatography.

• Research Organization: SYN-TEK AB, Umea (Sweden)
• OSTI ID: 7143888
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:19; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

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59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ABSORPTION SPECTROSCOPY
AMINO ACID SEQUENCE
ANIMAL CELLS
ANIMALS
ANTIBODIES
CELL CONSTITUENTS
CHO CELLS
COPPER COMPOUNDS
DNA
ELECTROPHORESIS
ELEMENTS
ENZYME IMMUNOASSAY
ENZYMES
FETAL MEMBRANES
IMMUNOASSAY
MAMMALS
MAN
MEMBRANES
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NONMETALS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
OXYGEN
PLACENTA
PLASMIDS
PRIMATES
PROTEINS
RECOMBINANT DNA
SECRETION
SPECTROSCOPY
SUPEROXIDE DISMUTASE
TRANSITION ELEMENT COMPOUNDS
VERTEBRATES
ZINC COMPOUNDS