Dihydroxypropylation of amino groups of proteins: use of glyceraldehyde as a reversible agent for reductive alkylation.
Dihydroxypropylation of amino groups of proteins: use of glyceraldehyde as a reversible agent for reductive alkylation.
复制标题
蛋白质氨基的二羟丙基化：使用甘油醛作为还原烷基化的可逆试剂。
DOI：
10.1021/bi00386a041
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Manjula,BN
中科院分区：
文献类型：
--
作者：
Acharya,AS;Manjula,BN
The Rockefeller University, New York, New York 10021 Received June 5, 1986; Revised Manuscript Received February 9, 1987 abstract: The mode of derivatization of amino groups of proteins by glyceraldehyde, an aldotriose, depends on the presence or absence of reducing agent. In the presence of sodium cyanoborohydride, the Schiff base adducts of the aldehyde with the amino groups are reduced, and dihydroxypropylation of amino groups takes place (reductive mode). The reductively glycated lysine residue, Art-(2, 3-dihydroxypropyl) lysine, is a substituted-amino alcohol. This-amino alcoholic function of the derivatized lysine should be susceptible to periodate oxidation, and thisoxidation is anticipated to result in the regeneration of the lysine residue. This aspect has been now investigated. Indeed, on mild periodate oxidation (15 mM periodate, 15 min at room temperature) of dihydroxypropylated ribonuclease A, nearly 95% of its Art-(2, 3-dihydroxypropyl) lysine residues were regenerated to lysine residues. The removal of the dihydroxypropyl groups by periodate oxidation could be accomplished within a wide pH range with little variation inthe recovery of lysines. The possible usefulness of this reversible chemical modification procedure in the primary structural studies of proteins was investigated with a tryptic peptide of dihydroxypropylated streptococcal M5 protein, namely, DHP-T4. This 12-residue tryptic peptide contains one internal iVf-(dihydroxypropyl) lysine. The di-hydroxypropylated peptidereleased most of its dihydroxypropyl groups on mild periodate oxidation. Redigestion of the periodate-treated peptide with trypsin generated the two expected peptides, demonstrating the generation of a trypsin-susceptible site. Reductive dihydroxypropylation of amino groups of RNase A resulted in the loss of its enzyme activity, the extent of inactivation increasing with the concentration of the glyceraldehyde used. Periodate oxidation of dihydroxypropylated ribonuclease A regenerated the full enzymic activity of the parent protein. These results demonstrate that dihydroxypropylation of amino groups is a valuable addition to the arsenal of reversible amino group modification procedures.
