Modification of hemoglobin with site-directed bifunctional reagents.
Modification of hemoglobin with site-directed bifunctional reagents.
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用定点双功能试剂修饰血红蛋白。
DOI：
10.1159/000205854
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发表时间：
1987
影响因子：
2.4
通讯作者：
Jones,RT
中科院分区：
文献类型：
--
作者：
Kavanaugh,MP;Shih,DT;Jones,RT
脱氧hb与烟酰胺腺嘌呤二核苷酸（o-NAD）、磷酸核糖基焦磷酸（o-PRPP）、三磷酸腺苷（o-ATP）、葡萄糖-1-磷酸（o-glc-1-P）和烟酰胺腺嘌呤二核苷酸磷酸（o-NADP）的高碘酸氧化衍生物反应，形成交联加合物，通过sds -聚丙烯酰胺凝胶电泳测定其产率不同。对Hb与o-NAD、o-PRPP和o-ATP交联进行氧平衡研究。这些衍生物被发现具有更高的氧亲和性，并在β链之间交联。六磷酸肌醇（IHP）阻断了这些试剂的修饰，表明修饰发生在有机磷结合位点。此外，研究发现双功能试剂4,4 ' -二异硫氰酸二苯乙烯-2,2 ' -二磺酸盐（DIDS）也能在β链之间形成交联的Hb，但其衍生物的氧亲和力显著降低，这使其成为基于Hb的无细胞血液替代品的潜在候选物
Reaction of deoxy-Hb with the periodate-oxidized derivatives of nicotinamide adenine dinucleotide (o-NAD), phosphoribosyl pyrophosphate (o-PRPP), adenosine triphosphate (o-ATP), glucose-1-phosphate (o-glc-1-P) and nicotinamide adenine dinucleotide phosphate (o-NADP) led to formation of cross-link adducts in varying yields as determined by SDS-polyacrylamide gel electrophoresis. Oxygen equilibrium studies were performed on Hb’s cross-linked with o-NAD, o-PRPP and o-ATP. These derivatives were found to have increased oxygen affinity and were cross-linked between theβchains. Inositol hexaphosphate (IHP) blocked modification by these reagents, suggesting that modification was occurring in the organic phosphate binding site. In addition, it was found that the bifunctional reagent 4,4’-diisothiocyanatostilbene-2,2’-disulfo-nate (DIDS) also led to formation of Hb cross-linked between theβchains, but resulted in a derivative with a dramatically decreased oxygen affinity, properties making it a potential candidate as an Hb-based cell-free blood substitute