Role of amino acid residues at the interface of alpha52asparginyl-N-glycosyl chain of human choriogonadotropin
| Autor: | Sathyamangalam V. Balasubramanian, Kui Shao, Om P. Bahl |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Circular dichroism
Protein Conformation Protein subunit Mutant Oligosaccharides Biology medicine.disease_cause Biochemistry Chorionic Gonadotropin Cell Line chemistry.chemical_compound Endocrinology medicine Animals Humans Glycosyl Amino Acids Molecular Biology chemistry.chemical_classification Mutation Circular Dichroism Wild type Biological activity Amino acid chemistry Asparagine |
| Zdroj: | Molecular and cellular endocrinology. 150(1-2) |
| ISSN: | 0303-7207 |
| Popis: | Of all the four N-glycosyl chains present in hCG, only one of them at alpha52Asn is located at the alpha/beta subunit interface and is crucial for the biological function of the hormone. The other three are exposed on the surface of the molecule and play only a minor role in the function of the hormone. The alpha52Asn oligosaccharide interacts with five amino acid residues in the beta-subunit, Tyr59, Val62, Phe64, Ala83, and Thr97. The present studies were undertaken to determine the role of the residues at the alpha52Asn-oligosaccharide and the beta-subunit interface in the mechanism of subunit association and downstream signaling events. Ten mutants, two of the alpha-subunit by the replacement of Asn52 and Thr80 with Gln and eight of the beta-subunit by multiple or single amino acid mutations, were prepared. These mutants included, hCGbeta59,62,64,97Ala, hCGbeta59,62,64Ala, hCGbeta62,64Ala, hCGbeta59Phe, hCGbeta62Ala or Thr, hCGbeta83Ile and hCGbeta97Ala. The mutation of the Asn52 to Gln resulted in a drastic change in its conformation and as a consequence in its weak affinity with the wild type beta as compared with that of the wild type hCGalpha and hCGalpha80Gln. The mutants with mutations in the four or three amino acids as well as both mutants of hCGbeta62Val almost failed to combine with hCGalpha again as a result of conformational changes shown by circular dichroism (CD) analysis and not due to their direct involvement in the subunit association. The double mutant combined with hCGalpha and the heterodimer behaved more like the wild type hCG. The mutation of Tyr to Phe resulted in a drop of 20% in the receptor binding and cAMP stimulation although Tyr is considered to be involved directly in subunit association. HCGbeta with mutations in the other amino acids, Phe64, Ala83, and Thr97, combined with the alpha subunit forming heterodimers with biological activity comparable to that of the wild type hCG. Thus, it appears that among the five amino acids in the vicinity of alpha52Asn carbohydrate, only beta59Tyr and beta62Val may be involved directly or indirectly in the alpha/beta beta dimer formation. |
| Databáze: | OpenAIRE |
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