STRUCTURE, PROPERTIES, AND FUNCTION OF UNSTABLE ANIMAL HEMOGLOBINS

本研究的目的是探讨心力衰竭的发病机制(S)。 血红蛋白结合/解离。它在体内的调节和 体外实验及其对血红蛋白功能的影响。软体动物， Anadara ovalis，其红细胞含有二聚体(HbMi)和 四聚体(HBMA)血红蛋白，两者均可解离为单体 在用K3Fe(CN)6氧化时(Borgese等人，1987年)。相比之下， 海生硬骨鱼，美洲鹅鱼，只有一只 形成聚集体的四聚体血红蛋白，其聚集度从160.000 当类似地氧化时超过900,000道尔顿(Borgese， 1986)。而许多种类的板鳃动物都有多个 血红蛋白，其中一些通过二硫键的形成聚合， 还没有关于海洋中的血红蛋白聚集的报道。 硬骨鱼。 我们的具体目标是：1)确定二硫化物的影响 还原剂对Hb的抑制和逆转作用 聚集(鹅鱼、海洋嘴唇)和单一化(阿纳达拉 Ovalis)。血红蛋白将通过DE-52层析和 分子量由凝胶过滤测定。2)我们会 确定是否可以通过以下方式在集成单元格中进行聚合 用NaN02等渗透性氧化剂孵育细胞，然后 填塞细胞的电子显微镜。3)完整的红细胞 有机磷酸盐分布将由Ag 1x8阴离子确定 交换层析；主要有机磷酸盐的影响 是否将使用Hem-O-Scan确定血红蛋白功能 氧离解分析仪。4)我们将确定细胞是否 具有代谢刺激的还原酶活性有更大的 抑制聚集的能力。5)我们将确定 Sh基团对鹅鱼和Anadara ovalis热稳定性的影响 HBMA。6)采用尿素凝胶电泳法和高效液相色谱仪 确定鹅类血红蛋白是同源还是异源四聚体。 分离链的氨基酸组成及测序 这些不寻常的血红蛋白(16SH/四聚体-鹅鱼和单体 Anadara Hb的形成)是解释结构的进一步步骤 真-功能-属性关系。 除了提供对表观Hb的解释之外 正常第四系血红蛋白的多态和改变 通过聚合或解离而形成的结构可能会严重 如果细胞形态和/或氧亲和力 受到不利影响。

The purpose of this research is to investigate the mechanism(s) of hemoglobin association/dissociation. Its regulation in vivo and in vitro and its effect on hemoglobin function. The mollusc, Anadara ovalis, has erythrocytes which contain dimeric (HbMi) and tetrameric (HbMa) hemoglobins both of which dissociate to monomers upon oxidation with K3Fe(CN)6 (Borgese et al., 1987). In contrast, the marine teleost, Lophius amercanus (goosefish), has a single tetrameric hemoglobin which forms aggregates ranging from 160.000 to greater than 900,000 daltons when similarly oxidized (Borgese, 1986). While many species of elasmobranchs have multiple hemoglobins, some of which polymerize by disulfide bond formation, there have been no reports of hemoglobin aggregation among marine teleosts. Our specific aims are: 1) to determine the effects of disulfide reducing agents on the inhibition and reversal of the Hb aggregation (goosefish, ocean pout) and monomerization (Anadara ovalis). Hemoglobins will be isolated by DE-52 chromatography and molecular weights determined by gel filtration. 2) We will determine whether aggregation can occur in intacts cells by incubating cells with permeable oxidants such as NaN02 followed by electron microscopy of the packed cells. 3) The complete red cell organic phosphate profile will be determined by Ag 1x8 anion exchange chromatography; the effect of the major organic phosphates are hemoglobin function will be determined using the Hem-O-Scan oxygen dissociation analyzer. 4) We will determine whether cells with metabolically stimulated reductase activity have a greater capacity to inhibit aggregation. 5) We will determine the role of SH groups on thermal stability of goosefish and Anadara ovalis HbMa. 6) Urea gel electrophoresis and HPLC will be used to determine whether goosefish hemoglobin is a homo or heterotetramer. Amino acid composition of the isolated chains and the sequencing of these unusual hemoglobins (16 SH/tetramer-goosefish and monomer formation of Anadara HBs) are further steps in explaining struc- ture-function-property relationships. In addition to providing an explanation for apparent Hb polymorphism, alterations in the normal quaternary hemoglobin structure by polymerization or dissociation may have serious consequences if cell morphology and/or oxygen affinity are adversely affected.