Synthesis of a hemoglobin polymer containing antioxidant enzymes using complementary chemistry of maleimides and sulfhydryls.
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.
Tarasov
Eugene
E
Department of Chemistry, Loyola University Chicago, Chicago, IL, USA.
Blaszak
Melanie M
MM
LaMarre
Jacqueline M
JM
Olsen
Kenneth W
KW
D001802:Blood Substitutes / Q000138:chemical synthesis* / Q000737:chemistry; D002374:Catalase / Q000737:chemistry; D006454:Hemoglobins / Q000138:chemical synthesis* / Q000737:chemistry; D006801:Humans; D008301:Maleimides / Q000737:chemistry*; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry*; D011108:Polymers / Q000138:chemical synthesis / Q000627:therapeutic use; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013482:Superoxide Dismutase / Q000737:chemistry
D001802:Blood Substitutes; D006454:Hemoglobins; D008301:Maleimides; D011108:Polymers; D013438:Sulfhydryl Compounds; C079129:hemoglobin polymer; D010088:Oxidoreductases; D002374:Catalase; D013482:Superoxide Dismutase