Selenoprotein W accumulates primarily in primate skeletal muscle, heart, brain and tongue

The human selenoprotein W coding region with the selenocysteine codon (TGA) changed to a cysteine codon (TGT) was fused to six histidine codons (at its 3' end), cloned into a prokaryotic expression vector (pTrc99a), and the corresponding mutated selenoprotein W was expressed in bacteria. The pr...

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Bibliographic Details
Published in:Molecular and cellular biochemistry 2000-01, Vol.204 (1-2), p.49-56
Main Authors: Gu, Q P, Sun, Y, Ream, L W, Whanger, P D
Format: Article
Language:English
Subjects:
Animals
Antibodies - immunology
Brain - metabolism
Escherichia coli
Glutathione Peroxidase - metabolism
Humans
In Vitro Techniques
Macaca mulatta
Muscle, Skeletal - metabolism
Muscular system
Mutation
Myocardium - metabolism
Organ Specificity
Primates
Proteins
Proteins - genetics
Proteins - immunology
Proteins - metabolism
Rabbits
RNA, Messenger - metabolism
Selenium
Selenium - metabolism
Selenoprotein W
Selenoproteins
Tongue - metabolism
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Summary:The human selenoprotein W coding region with the selenocysteine codon (TGA) changed to a cysteine codon (TGT) was fused to six histidine codons (at its 3' end), cloned into a prokaryotic expression vector (pTrc99a), and the corresponding mutated selenoprotein W was expressed in bacteria. The protein was purified by Ni-NTA agarose column and reverse phase HPLC. Polyclonal antibodies raised against this protein were used in Western blots to determine tissue distribution of selenoprotein W from rhesus monkeys fed a commercial chow. Selenoprotein W was found in several tissues with highest amounts in skeletal muscle and heart (muscle 6 fold greater than liver) and lowest levels in liver, but selenium concentrations were highest in kidneys (10 fold greater than muscle) and lowest in skeletal muscle. Northern blots using a human selenoprotein W cDNA probe indicated that mRNA levels were highest in monkey skeletal muscle and heart (2-2.5 fold greater than in liver), which is similar to the pattern found with a human multiple tissue Northern blot. However, as in the monkey, selenium concentrations were highest in human kidney and lowest in skeletal muscle and heart. Thus, selenoprotein W protein levels correlated with selenoprotein W mRNA levels but not with tissue selenium concentrations.
ISSN:0300-8177
1573-4919
DOI:10.1023/A:1007065829068