title: Keyhole limpet hemocyanin: structural and functional characterization of two different subunits and multimers.
abstract: 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.
authors.affiliation: PerImmune, Inc., Rockville, MD, USA.
authors.forename: R D
authors.initials: RD
authors.lastname: Swerdlow
authors.forename: R F
authors.initials: RF
authors.lastname: Ebert
authors.forename: P
authors.initials: P
authors.lastname: Lee
authors.forename: C
authors.initials: C
authors.lastname: Bonaventura
authors.forename: K I
authors.initials: KI
authors.lastname: Miller
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006433:Hemocyanins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D006801:Humans; D007167:Immunotherapy; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D008974:Mollusca; D010449:Peptide Mapping; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D001749:Urinary Bladder Neoplasms / Q000628:therapy
chemical_list: D046911:Macromolecular Substances; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin