Thiolation of chitosan. Attachment of proteins via thioether formation.
Thiolation of chitosan. Attachment of proteins via thioether formation.
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DOI：
10.1021/bm049352e
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发表时间：
2005-02
影响因子：
6.2
通讯作者：
T. Masuko;A. Minami;N. Iwasaki;T. Majima;S. Nishimura;Yuan-chuan Lee
中科院分区：
文献类型：
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作者：
T. Masuko;A. Minami;N. Iwasaki;T. Majima;S. Nishimura;Yuan-chuan Lee
壳聚糖通过将其他化合物与其氨基和羟基偶联，具有多种生物学功能。为了进一步扩大壳聚糖的应用范围，我们用2-亚胺基硫杂环戊烷对壳聚糖的氨基进行了修饰，得到了约20%的产率，相当于913微当量的SH/g壳聚糖或457nequiv SH/nmol的壳聚糖。将牛血清白蛋白与马来酰亚胺酯(N-(epsilon-maleimidocaproyloxy)sulfosuccinimide-EMCS)反应，得到马来酰亚胺修饰的牛血清白蛋白(MalN-BSA)。磺基EMCS加成的产率为12.8~36.8molMalN/molBSA。当壳聚糖-SH通过硫醚与MalN-BSA反应时，97.8%的马来酰亚胺基团被反应，37.2%的SH基团被消耗。其余SH组用溴乙酰胺猝灭。这是首次报道蛋白质与壳聚糖的共价偶联。我们的方法在开发新的壳聚糖生物医学材料方面应该会有很多应用，这些材料含有其他成分，如生长因子和细胞黏附分子，这些已知对细胞至关重要。我们的硫代化壳聚糖将促进这些生物医学成分的结合，为组织工程提供新型材料。
Chitosan has a variety of biological functions through conjugating of other compounds to their amino and hydroxyl groups. To further expand applicability of chitosan, we have modified the amino group of chitosan with 2-iminothiolane to bestow thiol groups and obtained about 20% yield, which is equivalent to 913 microequiv SH/g chitosan or 457 nequiv SH/nmol chitosan. Bovine serum albumin (BSA) was reacted with N-(epsilon-maleimidocaproyloxy)sulfosuccinimide ester (sulfo-EMCS), and maleimide-modified BSA (MalN-BSA) was obtained. The yield of sulfo-EMCS addition was 12.8-36.8 mol MalN/mol BSA. When the chitosan-SH was reacted with MalN-BSA via thioether, 97.8% of the maleimide group was reacted, and 37.2% of the SH group was consumed. The remaining SH group was quenched by bromoacetamide. This is the first report of covalent conjugation of a protein to chitosan. Our method should find many applications in developing new chitosan-based biomedical materials containing other components such as growth factors and cell adhesion molecules, known to be crucial to cells. Our thiolated chitosan will facilitate conjugation of such biomedical components to provide new types of materials for tissue engineering.