δ‐Aminolevulinic acid biosynthesis in <i>Ustilago maydis</i>

Schneegurt, Mark

doi:10.1002/jobm.200410479

Cited by 4 publications

(3 citation statements)

References 17 publications

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“…In living organisms there are two possible routes that have been reported for ALA synthesis. 76 In the mitochondria of animals, yeast and fungi, ALA results from condensation of succinyl CoA and glycine, catalysed by ALA synthetase. 77 Plants, algae and nearly all bacterial groups utilise the 5-carbon route to biosynthesis, where glutamate is converted to ALA along a three enzyme pathway.…”

Section: Synthesis Of Ala and Its Derivativesmentioning

confidence: 99%

Derivatives of 5-Aminolevulinic Acid for Photodynamic Therapy

Donnelly

McCarron

Woolfson

2007

Perspect Medicin Chem

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Photodynamic therapy (PDT) is a clinical treatment that combines the effects of visible light irradiation with subsequent biochemical events that arise from the presence of a photosensitising drug (possessing no dark toxicity) to cause destruction of selected cells. Today, the most common agent used in dermatological PDT is 5-aminolevulinic acid (ALA). As a result of its hydrophilic character, ALA penetrates skin lesions poorly when applied topically. Its systemic bioavailability is limited and it is known to cause significant side effects when given orally or intravenously. Numerous chemical derivatives of ALA have been synthesised with the aims of either improving topical penetration or enhancing systemic bioavailability, while reducing side effects. In vitro cell culture experiments with ALA derivatives have yielded promising results. However, if ALA derivatives are to demonstrate meaningful clinical benefits, a rational approach to topical formulation design is required, along with a systematic study aimed at uncovering the true potential of ALA derivatives in photodynamic therapy. With respect to systemic ALA delivery, more study is required in the developing area of ALA-containing dendrons and dendrimers.

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Section: Synthesis Of Ala and Its Derivativesmentioning

confidence: 99%

Derivatives of 5-Aminolevulinic Acid for Photodynamic Therapy

Donnelly

McCarron

Woolfson

2007

Perspect Medicin Chem

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“…Whilst many orthologues of genes predicted to encode ALA synthases have been described from various fungal genome sequencing projects, only enzymes from the yeast Saccharomyces cerevisiae and the basidiomycete Ustilago maydis have been scrutinised in detail (Schneegurt, 2005;Urban-Grimal & Labbe-Bois, 1981;Urban-Grimal et al, 1986). The gene encoding ALA synthase in Aspergillus oryzae (HemA) has been disrupted for the purpose of its development as a selectable marker in subsequent transformation (Elrod et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

δ-Aminolaevulinic acid synthesis is required for virulence of the wheat pathogen Stagonospora nodorum

2006

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d-Aminolaevulinic acid (ALA) is synthesized in fungi by ALA synthase, a key enzyme in the synthesis of haem. The requirement for ALA synthase in Stagonospora nodorum to cause disease in wheat was investigated. The single gene encoding ALA synthase (Als1) was cloned and characterized. Expression analysis determined that Als1 transcription was up-regulated during germination and also towards the latter stages of the infection. The Als1 gene was further characterized by homologous gene replacement. The inactivation of Als1 resulted in strains producing severely stunted germ tubes leading quickly to death. The strains could be recovered by supplementation with 33 mM ALA. Pathogenicity assays revealed the als1 strains were essentially non-pathogenic, inferring a key role for the synthesis of ALA during in planta growth. Supplementing the strains with ALA restored growth in vitro and also pathogenicity for up to 5 days after inoculation. Further examination by inoculating the als1 strains onto wounded leaves found that pathogenicity was only partially restored, suggesting that host-derived in planta levels of ALA are not sufficient to support growth. This study has identified a key role for fungal ALA synthesis during infection and revealed its potential as an antifungal target.

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“…Others have used coupled assays to enzymatically convert ALA to PBG [16]. Radiolabeled glycine has also been employed to generate labeled ALA [17]. ALA synthases are mitochondrial enzymes and some methods to assay activity have necessitated isolating mitochondrial fractions from tissues of interest in order to maximize the amount of ALA generated.…”

Section: Introductionmentioning

confidence: 99%

A method for determining δ-aminolevulinic acid synthase activity in homogenized cells and tissues

Bergonia¹,

Franklin²,

Kushner³

et al. 2015

Clinical Biochemistry

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Objective In mammalian cells the rate-limiting step in heme biosynthesis is the formation of δ-aminolevulinic acid (ALA). The reaction intermediates, porphyrins and iron and the final product, heme can be highly cytotoxic if allowed to accumulate. The importance of maintaining the levels of metabolic intermediates and heme within a narrow range is apparent based on the complex homeostatic system(s) that have developed. Ultimately, determining the enzymatic activity of ALA synthase (ALAS) present in the mitochondria is highly beneficial to confirm the effects of the transcriptional, translational and post-translational events. The aim of this study was to develop a highly sensitive assay for ALAS that could be used on whole tissue or cellular homogenates. Design and methods A systematic approach was used to optimize steps in formation of ALA by ALAS. Reducing the signal to noise ratio for the assay was achieved by derivatizing the ALA formed into a fluorescent product that could be efficiently separated by ultra performance liquid chromatography (UPLC) from other derivatized primary amines. The stability of whole tissue homogenate and cellular homogenate was determined after extended storage at −80°C. Conclusions A method for assaying ALAS has been developed that can be used with tissue homogenates or cellular lysates. There is no need to purify mitochondria and radiolabeled substrates are not needed for this assay. General laboratory reagents can be used to prepare the samples. Standard UPLC chromatography will resolve the derivatized ALA peak. Samples of tissue homogenate can be stored for approximately one year without significant loss of enzymatic activity.

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δ‐Aminolevulinic acid biosynthesis in Ustilago maydis

Cited by 4 publications

References 17 publications

Derivatives of 5-Aminolevulinic Acid for Photodynamic Therapy

Derivatives of 5-Aminolevulinic Acid for Photodynamic Therapy

δ-Aminolaevulinic acid synthesis is required for virulence of the wheat pathogen Stagonospora nodorum

A method for determining δ-aminolevulinic acid synthase activity in homogenized cells and tissues

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