USE OF MALEIC ANHYDRIDE FOR REVERSIBLE BLOCKING OF AMINO GROUPS IN POLYPEPTIDE CHAINS
USE OF MALEIC ANHYDRIDE FOR REVERSIBLE BLOCKING OF AMINO GROUPS IN POLYPEPTIDE CHAINS
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DOI：
10.1042/bj1120679
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发表时间：
1969-01-01
影响因子：
4.1
通讯作者：
LEBERMAN, R
中科院分区：
文献类型：
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作者：
BUTLER, PJG;HARRIS, JI;LEBERMAN, R
1. 马来酸酐与蛋白质和多肽的氨基反应迅速而特异。乳糜蛋白酶原在温和的条件下被完全取代，反应的程度可以很容易地从马来酰蛋白的光谱中确定。2. 马来酰蛋白通常是可溶的，在中性ph下分解。胰蛋白酶只在精氨酸残基上分裂被阻断的蛋白质，并且这种分裂通常有选择性，例如在酵母醇脱氢酶和猪甘油醛3-磷酸脱氢酶中。3. 该基团在酸性ph下通过分子内催化去除，半衰期为11-12hr。37°，pH3·5，在γ -马来酰赖氨酸或γ -马来酰凝乳胰蛋白酶原中。4. 解封反应可以作为赖氨酸肽和n端肽的“对角”电泳分离的基础，如β-促黑素细胞激素的研究所示。
1. Maleic anhydride was shown to react rapidly and specifically with amino groups of proteins and peptides. Complete substitution of chymotrypsinogen was achieved under mild conditions and the extent of reaction could be readily determined from the spectrum of the maleyl-protein. 2. Maleyl-proteins are generally soluble and disaggregated at neutral pH. Trypsin splits the blocked proteins only at arginine residues and there is frequently selectivity in this cleavage, e.g. in yeast alcohol dehydrogenase and pig glyceraldehyde 3-phosphate dehydrogenase. 3. The group is removed by intramolecular catalysis at acid pH. The half-time was 11–12hr. at 37° at pH3·5 in ∈-maleyl-lysine or in maleyl-chymotrypsinogen. 4. The unblocking reaction can be used as the basis for a ‘diagonal’-electrophoretic separation of lysine peptides andN-terminal peptides, as shown by studies with β-melanocyte-stimulating hormone.