EurekaMag
+ Most Popular
Cunninghamia lanceolata plantations in China
Mammalian lairs in paleo ecological studies and palynology
Studies on technological possibilities in utilization of anhydrous milk fat for production of recombined butter-like products
Should right-sided fibroelastomas be operated upon?
Large esophageal lipoma
Apoptosis in the mammalian thymus during normal histogenesis and under various in vitro and in vivo experimental conditions
Poissons characoides nouveaux ou non signales de l'Ilha do Bananal, Bresil
Desensitizing efficacy of Colgate Sensitive Maximum Strength and Fresh Mint Sensodyne dentifrices
Administration of fluid by subcutaneous infusion: revival of a forgotten method
Tundra mosquito control - an impossible dream?
Schizophrenia for primary care providers: how to contribute to the care of a vulnerable patient population
Geochemical pattern analysis; method of describing the Southeastern limestone regional aquifer system
Incidence of low birth weights in a hospital of Mexico City
Tabanidae
Graded management intensity of grassland systems for enhancing floristic diversity
Microbiology and biochemistry of cheese and fermented milk
The ember tetra: a new pygmy characid tetra from the Rio das Mortes, Brazil, Hyphessobrycon amandae sp. n. (Pisces, Characoidei)
Risk factors of contrast-induced nephropathy in patients after coronary artery intervention
Renovation of onsite domestic wastewater in a poorly drained soil
Observations of the propagation velocity and formation mechanism of burst fractures caused by gunshot
Systolic blood pressure in a population of infants in the first year of life: the Brompton study
Haematological studies in rats fed with metanil yellow
Studies on pasteurellosis. I. A new species of Pasteurella encountered in chronic fowl cholera
Dormancy breaking and germination of Acacia salicina Lindl. seeds
therapy of lupus nephritis. a two-year prospective study

Prelytic and lytic conformations of erythrocyte-associated Escherichia coli hemolysin


Prelytic and lytic conformations of erythrocyte-associated Escherichia coli hemolysin



Infection and Immunity 65(6): 2233-2239



ISSN/ISBN: 0019-9567

PMID: 9169756

Flow cytometry was developed as a method to assess the conformation of erythrocyte-bound Escherichia coli hemolysin polypeptide (HlyA). Topology of membrane-associated hemolysin (HlyA(E)) was investigated by testing surface accessibility of HlyA regions in lytic and nonlytic bound states, using a panel of 12 anti-HlyA monoclonal antibodies (MAbs). Hemolysin associates nonlytically with erythrocytes at 0 to 2 degrees C. To test the hypothesis that the nonlytic HlyA(E) conformation at 0 to 2 degrees C differs from the lytic conformation at 23 degrees C, MAb epitope reactivity profiles at the two temperatures were compared by flow cytometry. Four MAbs have distinctly increased reactivity at 0 to 2 degrees C compared to 23 degrees C. HlyA requires HlyC-dependent acylation at lysine residues 563 and 689 for lytic function. Toxin with cysteine substitution mutations at each lysine (HlyA(K563C) and HlyA(K689C)) as well as the nonacylated form of hemolysin made in a HlyC-deficient strain were examined by flow cytometry at 0 to 2 and 23 degrees C. The three mutants bind erythrocytes at wild-type toxin levels, but there are conformational changes reflected by altered MAb epitope accessibility for six of the MAbs. To test further the surface accessibility of regions in the vicinity of MAb-reactive epitopes, HlyA(E) was proteolytically treated prior to testing for MAb reactivity. Differences in protease susceptibility at 0 to 2 degrees and 23 degrees C for the reactivities of three of the MAbs further support the model of two distinct conformations of cell-associated toxin.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 009232791

Download citation: RISBibTeXText

Related references

Characterization and modification of Escherichia coli hemolysin cysteine substitution mutants Identification of a conformational difference between pre-lytic and lytic states on erythrocytes. Zentralblatt fuer Bakteriologie Suppl 29: 273-274, 1998

Therapeutic applications of pore-forming lytic toxins: potential use of Escherichia coli alpha-hemolysin. Medicina 62(1): 66-72, 2002

Induction of erythrocyte microvesicles by Escherichia Coli Alpha hemolysin. Biochemical Journal 476(22): 3455-3473, 2019

Interaction of escherichia coli alpha hemolysin with erythrocyte membranes. Abstracts of the Annual Meeting of the American Society for Microbiology 88: 44, 1988

Effect of Escherichia coli hemolysin on permeability of erythrocyte membranes to calcium. Toxicon: Official Journal of the International Society on Toxinology 24(6): 559-566, 1986

Calcium is required for binding of Escherichia coli hemolysin (HlyA) to erythrocyte membranes. Infection and Immunity 58(6): 1951-1958, 1990

P2X receptor-dependent erythrocyte damage by α-hemolysin from Escherichia coli triggers phagocytosis by THP-1 cells. Toxins 5(3): 472-487, 2013

Possible role of phospholipids in the binding of escherichia coli alpha hemolysin to sheep erythrocyte membranes. Abstracts of the Annual Meeting of the American Society for Microbiology 87: 29, 1987

Domains of Escherichia coli hemolysin (HlyA) involved in binding of calcium and erythrocyte membranes. Infection and Immunity 58(6): 1959-1964, 1990

Inhibition of lytic activity of Escherichia coli toxin hemolysin E against human red blood cells by a leucine zipper peptide and understanding the underlying mechanism. Biochemistry 47(7): 2134-2142, 2008

Location of tryptophan residues in free and membrane bound Escherichia coli alpha-hemolysin and their role on the lytic membrane properties. Biochimica et Biophysica Acta 1464(1): 27-34, 2000

Salmonella typhimurium strains carrying haemolysin plasmids and cloned haemolysin genes from Escherichia coli. Annales de l'Institut Pasteur. Microbiologie 136a(3): 289-301, 1985

Complementation of transport-deficient mutants of Escherichia coli a-hemolysin by second-site mutations in the transporter hemolysin B. The Journal of Biological Chemistry 268: 889-95, 1993

Restriction Fragment Length Polymorphisms Associated with -Hemolysin Determinants are Correlating with the Expression of -Hemolysin in Strains of Escherichia coli. Zentralblatt für Bakteriologie 276(2): 152-164, 1992

Complementation of transport-deficient mutants of Escherichia coli alpha-hemolysin by second-site mutations in the transporter hemolysin B. Journal of Biological Chemistry 268(26): 19889-19895, 1993