Cloning, sequencing, and expression of cDNA for human. beta. -glucuronidase

Journal Article · Sun Feb 01 04:00:00 EST 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.3.685· OSTI ID:6022997

Oshima, A; Kyle, J W; Miller, R D; Hoffmann, J W; Powell, P P; Grubb, J H; Sly, W S; Tropak, M; Guise, K S; Gravel, R A

The authors report here the cDNA sequence for human placental ..beta..-glucuronidase (..beta..-D-glucuronoside glucuronosohydrolase, EC 3.2.1.31) and demonstrate expression of the human enzyme in transfected COS cells. They also sequenced a partial cDNA clone from human fibroblasts that contained a 153-base-pair deletion within the coding sequence and found a second type of cDNA clone from placenta that contained the same deletion. Nuclease S1 mapping studies demonstrated two types of mRNAs in human placenta that corresponded to the two types of cDNA clones isolated. The NH/sub 2/-terminal amino acid sequence determined for human spleen ..beta..-glucuronidase agreed with that inferred from the DNA sequence of the two placental clones, beginning at amino acid 23, suggesting a cleaved signal sequence of 22 amino acids. When transfected into COS cells, plasmids containing either placental clone expressed an immunoprecipitable protein that contained N-linked oligosaccharides as evidenced by sensitivity to endoglycosidase F. However, only transfection with the clone containing the 153-base-pair segment led to expression of human ..beta..-glucuronidase activity. These studies provide the sequence for the full-length cDNA for human ..beta..-glucuronidase, demonstrate the existence of two populations of mRNA for ..beta..-glucuronidase in human placenta, only one of which specifies a catalytically active enzyme, and illustrate the importance of expression studies in verifying that a cDNA is functionally full-length.

• Research Organization: St. Louis Univ. School of Medicine, MO
• OSTI ID: 6022997
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:3; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550601 -- Medicine-- Unsealed Radionuclides in Diagnostics
59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
AMINO ACID SEQUENCE
AMINO ACIDS
ANIMAL CELLS
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CLONING
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA
DNA SEQUENCING
DNA-CLONING
DRUGS
ENZYMES
EVEN-ODD NUCLEI
FETAL MEMBRANES
FIBROBLASTS
GENE REGULATION
GLUCURONIDASE
GLYCOSYL HYDROLASES
HYBRIDIZATION
HYDROLASES
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MAN
MEMBRANES
METHIONINE
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
O-GLYCOSYL HYDROLASES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLACENTA
PRIMATES
RADIOISOTOPES
RECOMBINANT DNA
RNA
SOMATIC CELLS
SPLEEN
STRUCTURAL CHEMICAL ANALYSIS
SULFUR 35
SULFUR ISOTOPES
VERTEBRATES