title: Structural studies on Laz, a promiscuous anticancer Neisserial protein.
abstract: Azurin and Laz (lipidated azurin) are 2 bacterial proteins with anticancer, anti-viral and anti-parasitic activities. Azurin, isolated from the bacterium Pseudomonas aeruginosa, termed Paz, demonstrates anticancer activity against a range of cancers but not against brain tumors. In contrast, Laz is produced by members of Gonococci/Meningococci, including Neisseria meningitides which can cross the blood-brain barrier to infect brain meninges. It has been previously reported that Laz has an additional 39 amino acid moiety, called an H.8 epitope, in the N-terminal part of the azurin moiety that allows Laz to cross the entry barrier to brain tumors such as glioblastomas. Exactly, how the H.8 epitope helps the azurin moiety of Laz to cross the entry barriers to attack glioblastoma cells is unknown. In this paper, we describe the structural features of the H.8 moiety in Laz using X-ray crystallography and demonstrate that while the azurin moiety of Laz adopts a β-sandwich fold with 2 β-sheets arranged in the Greek key motif, the H.8 epitope was present as a disordered structure outside the Greek key motif. Structures of Paz and H.8 epitope-deficient Laz are well superimposed. The structural flexibility of the H.8 motif in Laz explains the extracellular location of Laz in Neisseria where it can bind the key components of brain tumor cells to disrupt their tight junctions and allow entry of Laz inside the tumors to exert cytotoxicity.
authors.affiliation: a Laboratory of Basic and Applied Molecular Biotechnology; Graduate School of Agriculture ; Kyoto University ; Uji , Kyoto , Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Hashimoto
authors.forename: Akihito
authors.initials: A
authors.lastname: Ochiai
authors.forename: Chang Soo
authors.initials: CS
authors.lastname: Hong
authors.forename: Kousaku
authors.initials: K
authors.lastname: Murata
authors.forename: Ananda M
authors.initials: AM
authors.lastname: Chakrabarty
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D001400:Azurin / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000494:pharmacology*; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009343:Neisseria / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism; D019108:Tight Junctions / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D001400:Azurin
keywords: H.8 epitope; X-ray crystallography; azurin; disorder; laz; neisseria