Tarasov Eugene, Blaszak Melanie M, LaMarre Jacqueline M, Olsen Kenneth W
Department of Chemistry, Loyola University Chicago, Chicago, IL, USA.
Artif Cells Blood Substit Immobil Biotechnol. 2007;35(1):31-43. doi: 10.1080/10731190600974434.
To increase the overall size of hemoglobin (Hb), we developed a novel system of polymerization based on the complementary chemistry between sulfhydryls and maleimides. The maleimides were introduced onto the protein through N-(-maleimidobutyryloxy) succinimide, while the sulfhydryls were added using 2-iminothiolane hydrochloride (Trauts reagent). Resulting polymers showed SDS-PAGE bands with molecular weights as high as 96 kDa. Size exclusion chromatography has demonstrated species with molecular weight > 700 kDa. The flexibility of the sulfhydryl-maleimide chemistry has also allowed insertion of two antioxidant enzymes, catalase (Cat) and superoxide dismutase (SOD), into the Hb polymer. Cat was incorporated into the heavier fractions of the polymer, while SOD was found throughout the molecular weight range.
为了增大血红蛋白（Hb）的整体尺寸，我们基于巯基与马来酰亚胺之间的互补化学原理开发了一种新型聚合体系。通过N-（-马来酰亚胺丁酰氧基）琥珀酰亚胺将马来酰亚胺引入蛋白质中，同时使用盐酸2-亚氨基乙硫醇（特劳茨试剂）添加巯基。所得聚合物在SDS-PAGE电泳中显示出分子量高达96 kDa的条带。尺寸排阻色谱法已证明存在分子量> 700 kDa的物种。巯基-马来酰亚胺化学的灵活性还允许将两种抗氧化酶，过氧化氢酶（Cat）和超氧化物歧化酶（SOD）插入到Hb聚合物中。Cat被并入聚合物的较重部分，而SOD在整个分子量范围内均有发现。
