Purification of cysteine-rich bioactive peptides from leukocytes by continuous acid-urea-polyacrylamide gel electrophoresis.
Purification of cysteine-rich bioactive peptides from leukocytes by continuous acid-urea-polyacrylamide gel electrophoresis.
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通过连续酸性-尿素-聚丙烯酰胺凝胶电泳从白细胞中纯化富含半胱氨酸的生物活性肽。
DOI：
10.1006/abio.1993.1065
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发表时间：
1993
影响因子：
2.9
通讯作者：
Lehrer,RI
中科院分区：
文献类型：
--
作者：
Harwig,SS;Chen,NP;Park,AS;Lehrer,RI
建立了一种新的连续酸尿素-聚丙烯酰胺凝胶电泳法(CAU-PAGE)，并将其应用于兔和人防御素的纯化。利用它，我们鉴定了兔防御素NP-1和NP-2的两种翻译后修饰形式，并从富含白细胞的人白细胞中纯化了一种加工过的RANTES(β-intercrine)肽。CAU-PAGE显示兔防御素NP-5的回收率约为70%。回收的防御素没有N-末端修饰，其体外抗菌活性与用上述层析方法纯化的防御素相当。由于CAU-PAGE是在非还原条件下进行的，因此它特别适用于分子内二硫键阳离子多肽的纯化，如防御素和α或β-Intercrine。
A new continuous acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE) preparative method was developed and used to purify rabbit and human defensins. With it, we identified two post-translationally modified forms of rabbit defensins NP-1 and NP-2, and purified a processed RANTES (β-intercrine) peptide from leukophoresed human leukocytes. CAU-PAGE afforded approximately 70% recovery of rabbit defensin NP-5. The recovered defensins were not N-terminally modified, and their in vitro antimicrobial activity was equivalent to that of defensins purified by previously described chromatographic methods. Since CAU-PAGE is performed under nonreducing conditions, it should be especially useful for purifying cationic peptides with intramolecular disulfide bonds, such as defensins and α or β-intercrines.