贵州医科大学学报

1990, (02) 102-106

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应用HPLC分析一例HbJ-Bangkok
HpLC procedures and DABITC/pITC Micro-Double Coupling Method in the Gequence Analysis of Hemoglobin Variant——The structure analysis of Hb J-Bangkok

陈尚武;单可人;单拓生;张小蕾;
Chen Shangwu, et al Department of Biochemistry

摘要(Abstract):

本文报道了将高效液相色谱(HpLC)技术引入蛋白质结构的研究工作,配合4-N,N-二甲胺基偶氮苯-4-异硫氰酸酯/异硫氰酸苯酯(DABITC/pITC)微量双偶合法对一例快泳Hb变异体的一级结构分析成功表明该异常Hbe链的第56位甘氨酸被天冬氨酸替代,即为HbJ-Bangkok(β56(D7)Gly→Asp)。在异常血红蛋白一级结构的研究中,HpLC和DABITC-pITC双偶合法的联合应用较通常使用的指纹图谱法有许多突出的优点。
This paper reports the chemical structure of a fastmoving β-chain hemoglobin variant, Hb J-Bangkok,determined by HpLC procedures and DABITC/pITC microdouble coupling method. The propositus, a 28 years old farmer, was a heterozygote and belonged to Han nationality. The abnormal hemoglobin migrated in front of HbA on cellulose acetate membrane at pH 8. 6. The content of abnormal hemoglobin was 48.4%-54.5%. The separation of globin chains was performed by using column chromatography with CM23. The purified globin chain was digested with TpCK-trypsin, and the tryptic digests were sparated with reverse-phase HpLC. The composition of amino acids of hydrolyzed peptides was also analysed quantitatively with the same HpLC system. Abnormal peptide sequence analysis was performed with DABITC/pITC micro-double coupling method. The results indicated that the glycine residue 56 was substituted by an aspartic acid residue.

关键词(KeyWords): 异常血红蛋白;氨基酸序列;高压液相色谱法
hemoglobins;abnormal;amino dcid seguence;chromatography;high pcrformane liquid

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作者(Author): 陈尚武;单可人;单拓生;张小蕾;
Chen Shangwu, et al Department of Biochemistry

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DOI: 10.19367/j.cnki.1000-2707.1990.02.006

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