Purification and characterization of a membrane-associated 3,3',5-triiodo-L-thyronine binding protein from a human carcinoma cell line
A membrane-associated binding protein for 3,3',5-triiodo-L-thyronine (T3) was purified to apparent homogeneity from A431 human epidermoid carcinoma cells. A431 cells were specifically labeled with the N-bromoacetyl derivative of T3 labeled with SVI at the 3' position (BrAc( SVI)T3) and were extracted with 3-(3-(cholamidopropyl)dimethylammonio)-1-propanesulfonate (CHAPS), a zwitterionic detergent. The solubilized BrAc( SVI)T3-labeled protein was successively purified by chromatography on Sephadex G-200 and QAE-Sephadex followed by NaDodSO4/PAGE. Approximately 0.2 mg of purified protein was obtained from 2.5 x 10Z cells, which represents a 3000-fold purification. The membrane-associated T3 binding protein is an acidic protein with a pI of 5.1 and an apparent molecular mass of 55,000 daltons determined by NaDodSO4/PAGE. Polyclonal antibodies against the 55-kDa protein were prepared and used in indirect immunofluorescence to show that the 55-kDa protein was mainly found in the nuclear envelope and endoplasmic reticulum.
- Research Organization:
- National Institutes of Health (NIH), Bethesda, MD (United States)
- OSTI ID:
- 6156008
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:4, Issue 4
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
LABELLED COMPOUNDS
CONFIGURATION INTERACTION
TRIIODOTHYRONINE
TUMOR CELLS
CARCINOMAS
CELL MEMBRANES
ELECTROPHORESIS
FLUORESCENCE
IODINE 125
LIQUID COLUMN CHROMATOGRAPHY
METHIONINE
MONOCLONAL ANTIBODIES
PROTEINS
PURIFICATION
SULFUR 35
AMINO ACIDS
ANIMAL CELLS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DISEASES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
EVEN-ODD NUCLEI
HORMONES
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
LUMINESCENCE
MEMBRANES
NEOPLASMS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDE HORMONES
RADIOISOTOPES
SEPARATION PROCESSES
SULFUR ISOTOPES
THYROID HORMONES
550201* - Biochemistry- Tracer Techniques