Swerdlow R D, Ebert R F, Lee P, Bonaventura C, Miller K I
PerImmune, Inc., Rockville, MD, USA.
Comp Biochem Physiol B Biochem Mol Biol. 1996 Mar;113(3):537-48. doi: 10.1016/0305-0491(95)02091-8.
Keyhole limpet hemocyanin (KLH), the large respiratory glycoprotein from the primitive gastropod mollusc, Megathura crenulata, is a potent immunogen used classically as a carrier protein for haptens and more recently in human vaccines and for immunotherapy of bladder cancer. Two KLH isoforms were identified and isolated by high-performance anion exchange chromatography. Subsequent analyses disclosed that these isoforms--designated KLH-A and KLH-B--were composed of distinct subunits that differed in primary structure, molecular weight (KLH-A was 449,000 and KLH-B was 392,000), polymerization/reassociation characteristics, and O2-binding constants (KLH-A had a P50 of 7.32 and KLH-B had a P50 of 2.46). Both subunits appear to be composed of eight oxygen binding domains, and reassociate in solution only with like subunits. These results support the concept that structural and functional heterogeneity is a common feature of molluscan hemocyanins, and provide a rational basis for studying and optimizing the immunostimulatory properties of KLH.
钥孔血蓝蛋白（KLH）是一种来自原始腹足纲软体动物巨缘帽贝（Megathura crenulata）的大型呼吸糖蛋白，是一种有效的免疫原，传统上用作半抗原的载体蛋白，最近也用于人类疫苗和膀胱癌的免疫治疗。通过高效阴离子交换色谱法鉴定并分离出两种KLH亚型。随后的分析表明，这些亚型——命名为KLH-A和KLH-B——由不同的亚基组成，这些亚基在一级结构、分子量（KLH-A为449,000，KLH-B为392,000）、聚合/重缔合特性和氧结合常数（KLH-A的P50为7.32，KLH-B的P50为2.46）方面存在差异。两个亚基似乎都由八个氧结合结构域组成，并且在溶液中仅与相同的亚基重缔合。这些结果支持了结构和功能异质性是软体动物血蓝蛋白的共同特征这一概念，并为研究和优化KLH的免疫刺激特性提供了合理依据。
