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Identification of intramolecular crosslinks in bovine growth hormone after two-step modification with 1,5-difluoro-2,4-dinitrobenzene.
Ermácora, M R; Nowicki, C; Wolfenstein-Todel, C; Santomé, J A.
Afiliação
  • Ermácora MR; Institute of Biochemistry and Biophysical Chemistry (UBA-CONICET), Faculty of Pharmacy and Biochemistry, Buenos Aires, Argentina.
Int J Pept Protein Res ; 30(3): 423-30, 1987 Sep.
Article em En | MEDLINE | ID: mdl-3692688
Derivatives of bovine growth hormone, containing monoaminotyrosyl residues in positions 35, 42 and 174, were treated at pH 3.6 with a bifunctional reagent, 1,5-difluoro-2,4-dinitrobenzene. Under these conditions aminotyrosyl groups reacted. On changing the pH to 9.3, the second fluorine atom of the reagent was substituted with the sterically adjacent side groups of lysine, since the excess of reagent had been previously removed. The modified protein underwent cyanogen bromide treatment. Peptides containing the crosslinks were purified from tryptic digests of the cyanogen bromide fragments by HPLC. Results show that aminoTyr 174 was able to form dinitrophenylene bridges with Lys 111, Lys 29 and Lys 170. AminoTry 35 was found crosslinked to Lys 29. Taking into account the size of the reagent, it may be inferred that Lys 29, 111 and 170 are located at approximately 5 A from Tyr 174 in the bovine growth hormone molecule.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Hormônio do Crescimento / Dinitrofluorbenzeno / Nitrobenzenos Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Int J Pept Protein Res Ano de publicação: 1987 Tipo de documento: Article País de afiliação: Argentina País de publicação: Dinamarca
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Hormônio do Crescimento / Dinitrofluorbenzeno / Nitrobenzenos Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Revista: Int J Pept Protein Res Ano de publicação: 1987 Tipo de documento: Article País de afiliação: Argentina País de publicação: Dinamarca