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Differential Sialic Acid Content and Hemoglobin-binding Activity of Precursor Prohaptoglobin and Mature Haptoglobin

전구체 프로합토글로빈과 성숙 합토글로빈의 시알산 함량 및 헤모글로빈-결합력 비교

  • Lee, Joo-Hyun (Department of Medical Lifescience, College of Medicine, The Catholic University of Korea) ;
  • Oh, Mi-Kyung (Department of Medical Lifescience, College of Medicine, The Catholic University of Korea) ;
  • Kim, In-Sook (Department of Medical Lifescience, College of Medicine, The Catholic University of Korea)
  • 이주현 (가톨릭대학교 의과대학 의생명과학교실) ;
  • 오미경 (가톨릭대학교 의과대학 의생명과학교실) ;
  • 김인숙 (가톨릭대학교 의과대학 의생명과학교실)
  • Received : 2017.04.07
  • Accepted : 2017.05.11
  • Published : 2017.06.30

Abstract

Mature haptoglobin (Hp) is a plasma glycoprotein and acts as an antioxidant by scavenging cell-free hemoglobin (Hb). Prohaptoglobin (proHp) is an unprocessed Hp precursor which is present a little in circulation. However, the biological function of proHp remains unknown. To investigate the structural and functional differences between proHp and Hp, we prepared recombinant proHp isoforms and compared their sialic acid content and Hb-binding capacity with those of mature isoforms. When proHp samples were analyzed by Western blot under non-reducing conditions, proHp1 was detected as one band of approximately 130 kDa and proHp2 as multiple bands >200 kDa, in the manner of mature Hp1-1 and Hp2-2, respectively. On the native polyacrylamide gel under non-reducing and non-denaturing conditions, both proHp isoforms migrated more slowly than their mature Hp counterparts. In addition, the lectin-based ELISA assay demonstrated that the content of sialic acid in proHp1 and proHp2 was much less than in Hp1-1 and Hp2-2. The Hb-binding capacity of proHp was also lower than those of mature Hp. These findings indicate that proHp and Hp are similar in the size and polymerization pattern, but different in sialic acid content and Hb-binding activity. It suggests precursor proHp may exert different functions in circulation than does mature Hp.

Acknowledgement

Supported by : NRF

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