Yamamoto T, Cochrane C G
Am J Pathol. 1981 Nov;105(2):164-75.
Hageman factor was purified from guinea pig plasma by successive column chromatography. The guinea pig Hageman factor appeared homogeneous as a single-chain protein on polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS) and beta-mercaptoethanol. The apparent molecular weight was 76,000 daltons by SDS--polyacrylamide gel electrophoresis and 105,000 daltons by gel filtration with a Sephadex G-150 column. Amino acid composition of the guinea pig Hageman factor was similar to that reported for human, bovine, and rabbit Hageman factors. The purified guinea pig Hageman factor, as well as guinea pig plasma, showed strong clotting time correction activity in Hageman-factor--deficient human plasma. The activity could be blocked by the IgG fraction of antiserums against guinea pig Hageman factor raised in rabbits or a goat. The concentration of Hageman factor in guinea pig plasma was determined to be 120 microgram/ml by quantitative radial immunodiffusion assay. The 28,000-dalton active form of Hageman factor (beta-HFa) was prepared from guinea pig Hageman factor by treatment with plasma kallikrein. beta-HFa caused an increase in vascular permeability when injected into guinea pig skin at concentrations as low as 3 x 10(-10) M (0.8 ng). Native, or zymogen Hageman factor did not cause an increase in permeability at concentrations of up to 2 x 10(-7) M. The increased permeability induced by beta-HFa was short lasting, with about a 50% decrease in activity apparent within 6 minutes after intradermal injection. The permeability enhancement activity of beta-HFa was inhibited by pretreatment of beta-HFa with diisopropylfluorophosphate. It may be concluded that active Hageman factor in the interstitial space of guinea pigs acts as a vascular permeability factor of far greater potency than bradykinin.
通过连续柱色谱法从豚鼠血浆中纯化了哈格曼因子。在十二烷基硫酸钠（SDS）和β-巯基乙醇存在的情况下，豚鼠哈格曼因子在聚丙烯酰胺凝胶上呈现为单链蛋白的均一状态。通过SDS-聚丙烯酰胺凝胶电泳，其表观分子量为76,000道尔顿，而用Sephadex G-150柱进行凝胶过滤时，表观分子量为105,000道尔顿。豚鼠哈格曼因子的氨基酸组成与报道的人、牛和兔哈格曼因子的氨基酸组成相似。纯化的豚鼠哈格曼因子以及豚鼠血浆在哈格曼因子缺乏的人血浆中显示出强烈的凝血时间校正活性。该活性可被兔或山羊体内产生的抗豚鼠哈格曼因子抗血清的IgG组分所阻断。通过定量放射免疫扩散测定法测定豚鼠血浆中哈格曼因子的浓度为120微克/毫升。通过用血浆激肽释放酶处理，从豚鼠哈格曼因子制备了28,000道尔顿的活性形式的哈格曼因子（β-HFa）。当以低至3×10⁻¹⁰ M（0.8纳克）的浓度注射到豚鼠皮肤中时，β-HFa会导致血管通透性增加。天然的或酶原性哈格曼因子在浓度高达2×10⁻⁷ M时不会导致通透性增加。β-HFa诱导的通透性增加持续时间较短，皮内注射后6分钟内活性明显下降约5０％。β-HFa的通透性增强活性可被用二异丙基氟磷酸对β-HFa进行预处理所抑制。可以得出结论，豚鼠间质空间中的活性哈格曼因子作为一种血管通透性因子，其效力远大于缓激肽。
