Abstract
The zoopathogenic fungus Histoplasma capsulatum, like other eukaryotic aerobic microorganisms, requires iron for growth. Under conditions of low iron availability, the fungus secretes hydroxamates that function as siderophores (iron chelators). The experiments to be reported were designed to gather further information on the hydroxamate siderophores of H. capsulatum. The fungus was grown in a synthetic medium deferrated with the cationic exchange resin Chelex 100. Siderophores were detected after 4 days of incubation at 37 degrees C in media containing 0.3 to 1.0 microM iron. The secretion was suppressed by 10 microM iron. The hydroxamates were purified by reverse-phase and size-exclusion chromatography. On the basis of ions observed during electrospray mass spectroscopy, five hydroxamate siderophores were tentatively identified: dimerum acid, acetyl dimerum acid, coprogen B, methyl coprogen B, and fusarinine (monomeric). A polyclonal antibody to dimerum acid was generated. This reagent cross-reacted with coprogen B and fusarinine. Thus, the antibody detects hydroxamates in all three families of siderophores excreted by H. capsulatum.
MeSH Terms
Antibodies
Chelating Agents/metabolism
Chromatography, High Pressure Liquid
Dose-Response Relationship, Drug
Histoplasma/immunology,metabolism
Hydrolysis
Hydroxamic Acids/chemistry,immunology,metabolism
Immunoblotting
Iron/metabolism
Mass Spectrometry
Piperazines/metabolism
Siderophores/chemistry,immunology,isolation & purification,metabolism
Time Factors
Chemicals
Antibodies
Chelating Agents
Hydroxamic Acids
Piperazines
Siderophores
coprogen B
fusarinine
dimerum acid
Iron
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Howard D H
Department of Microbiology, Neuropsychiatric Institute, UCLA School of Medicine, Los Angeles, California, USA. dhoward@ucla.edu
Rafie R
Tiwari A
Faull K F
References (18)
18 references, click to expand
-
Nutritional studies of Histoplasma capsulatum.
Mycopathol Mycol Appl. 1965 Apr 14;25(3):294-308
PMID: 5877181
-
Acquisition, transport, and storage of iron by pathogenic fungi.
Clin Microbiol Rev. 1999 Jul;12(3):394-404
PMID: 10398672
-
Low molecular weight intracellular iron transport compounds.
Blood. 1977 Sep;50(3):433-9
PMID: 328083
-
Enzymatic release of iron from sideramines in fungi. NADH:sideramine oxidoreductase in Neurospora crassa.
Biochim Biophys Acta. 1977 Nov 7;500(1):27-41
PMID: 144535
-
Identification of coprogen B and its breakdown products from Histoplasma capsulatum.
Infect Immun. 1982 Mar;35(3):990-6
PMID: 7068227
-
Isolation and characterization of spontaneous avirulent variants of Histoplasma capsulatum.
Infect Immun. 1987 Mar;55(3):528-33
PMID: 3818083
-
Chelation of transferrin iron by desferrioxamine in K562 cells. The partition of iron between ferrioxamine and ferritin.
Biochem J. 1988 Sep 15;254(3):869-75
PMID: 3196300
-
Mobilization of iron from endocytic vesicles. The effects of acidification and reduction.
J Biol Chem. 1990 Apr 25;265(12):6688-92
PMID: 2324097
-
High-performance liquid chromatography of siderophores from fungi.
Biol Met. 1988;1(1):9-17
PMID: 2978959
-
Iron limitation and the gamma interferon-mediated antihistoplasma state of murine macrophages.
Infect Immun. 1991 Jul;59(7):2274-8
PMID: 1904840
-
Characterization of the micro-environment of Salmonella typhimurium-containing vacuoles within MDCK epithelial cells.
Mol Microbiol. 1992 Nov;6(22):3289-97
PMID: 1484485
-
Chloroquine induces human macrophage killing of Histoplasma capsulatum by limiting the availability of intracellular iron and is therapeutic in a murine model of histoplasmosis.
J Clin Invest. 1994 Apr;93(4):1422-9
PMID: 8163646
-
Iron acquisition in microbial pathogenesis.
Trends Microbiol. 1993 May;1(2):66-9
PMID: 8044465
-
Inhibition of growth of Histoplasma capsulatum yeast cells in human macrophages by the iron chelator VUF 8514 and comparison of VUF 8514 with deferoxamine.
Antimicrob Agents Chemother. 1995 Aug;39(8):1824-9
PMID: 7486926
-
Synthesis of optically pure chrysobactin and immunoassay development.
Biometals. 1996 Oct;9(4):377-83
PMID: 8837459
-
Quorum sensing: the explanation of a curious phenomenon reveals a common characteristic of bacteria.
J Bacteriol. 1999 May;181(9):2667-8
PMID: 10217751
-
A symphony of bacterial voices.
Science. 1999 May 21;284(5418):1302-4
PMID: 10383312
-
Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine).
Anal Biochem. 1971 Apr;40(2):450-8
PMID: 5551554