Nowicki C, Wolfenstein-Todel C, Santomé J A
Int J Pept Protein Res. 1985 Dec;26(6):568-74. doi: 10.1111/j.1399-3011.1985.tb03213.x.
Bovine growth hormone was modified by reaction with 1,5-difluoro-2,4-dinitrobenzene under conditions favouring production of intramolecularly crosslinked derivatives from monomeric molecules. The monomeric fraction, isolated by chromatography on Sephadex G-100, was oxidized or reduced and carbamidomethylated and trypsin digested. The resulting peptides were fractionated on SP-Sephadex and further purified by peptide mapping or HPLC. Two modified peptides containing sequences 108-112 or 108-113 and 171-176 of bGH were obtained, including a dinitrophenylene bridge between lysine 111 and tyrosine 174, thus suggesting the stereochemical proximity of these residues.
牛生长激素在有利于从单体分子生成分子内交联衍生物的条件下与1,5-二氟-2,4-二硝基苯反应进行修饰。通过在Sephadex G-100上进行色谱分离得到单体部分,将其氧化或还原,进行氨基甲酰甲基化并经胰蛋白酶消化。所得肽在SP-Sephadex上进行分级分离,并通过肽图谱或高效液相色谱进一步纯化。获得了两条修饰肽,其包含bGH的108-112或108-113以及171-176序列,包括赖氨酸111和酪氨酸174之间的二亚硝基苯桥,从而表明这些残基在立体化学上的接近性。