The N-Acetyl-D-glucosaminylphosphatidylinositol De-N-acetylase of Glycosylphosphatidylinositol Biosynthesis Is a Zinc Metalloenzyme

Urbaniak, Michael D.; Crossman, Arthur; Chang, Tunhan; Smith, Terry; Aalten, Daan M. F. van; Ferguson, Michael A. J.

doi:10.1074/jbc.m502402200

Cited by 39 publications

(59 citation statements)

References 38 publications

(44 reference statements)

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“…More recent reports on the active domain from rat PIG-L have suggested that it is an extremely 1,10-PO-sensitive metalloenzyme that does not recover activity upon long incubation with the chelator (18). However, the E. histolytica PIG-L appears to be different from rat PIG-L because a similar sensitivity to either EDTA or 1,10-PO was not observed in our assays.…”

Section: Discussioncontrasting

confidence: 41%

“…Divalent Cations Stimulate De-N-acetylation-Previous reports have suggested that the catalytic domain of rat PIG-L is a metalloprotein that lost activity irreversibly when treated with metal chelators for long periods of time (8,18). Hence, we tested whether MBPEh⌬TMPIG-L was also a metal-dependent de-N-acetylase.…”

Section: Tm Deletion Results In a Functionally Folded Protein Capable Ofmentioning

confidence: 99%

“…Hence, the de-N-acetylase activity of this protein was assayed in the presence of bound GroEL in both the absence and the presence of detergent as in Ref. 18.…”

Section: Resultsmentioning

confidence: 99%

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N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase from Entamoeba histolytica

Ashraf¹,

Yadav²,

Sreejith

et al. 2011

Journal of Biological Chemistry

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PIG-L/GPI12 proteins are endoplasmic reticulum-resident membrane proteins involved in the second step of glycosylphosphatidylinositol anchor biosynthesis in eukaryotes. We show that the Entamoeba histolytica PIG-L protein is optimally active in the acidic pH range. The enzyme has an intrinsic low level of de-N-acetylase activity in the absence of metal and is significantly stimulated by divalent cations. Metal binding induces a large conformational change in the protein that appears to improve catalytic rates while not altering the affinity of the enzyme for its substrate.

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Section: Discussioncontrasting

confidence: 41%

Section: Tm Deletion Results In a Functionally Folded Protein Capable Ofmentioning

confidence: 99%

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N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase from Entamoeba histolytica

Ashraf¹,

Yadav²,

Sreejith

et al. 2011

Journal of Biological Chemistry

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“…This was somewhat surprising, because low-GC bacteria, including Staphylococcus, do not have MSH metabolism. However, MSH S-conjugate amidase and the other M. tuberculosis paralogs, MshB and the uncharacterized protein Rv0323c, belong to a large family of zinc metalloproteinases with homologs in eukaryotes (143,154), low-GC gram-positive bacteria, such as the staphylococci and the bacilli (25,139), and DeinococcusThermus (42). Since the bromotyrosine natural products (100) and analogs contain the zinc-binding hydroxamate moiety, it is not difficult to envision inhibition of this large family of proteins by inhibitors such as EXEG1706 (Fig.…”

Section: Msh Drug Targets In Mycobacterium Tuberculosismentioning

confidence: 99%

Biosynthesis and Functions of Mycothiol, the Unique Protective Thiol of Actinobacteria

Newton

Buchmeier

Fahey

2008

Microbiol Mol Biol Rev

313

293

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SUMMARY Mycothiol (MSH; AcCys-GlcN-Ins) is the major thiol found in Actinobacteria and has many of the functions of glutathione, which is the dominant thiol in other bacteria and eukaryotes but is absent in Actinobacteria. MSH functions as a protected reserve of cysteine and in the detoxification of alkylating agents, reactive oxygen and nitrogen species, and antibiotics. MSH also acts as a thiol buffer which is important in maintaining the highly reducing environment within the cell and protecting against disulfide stress. The pathway of MSH biosynthesis involves production of GlcNAc-Ins-P by MSH glycosyltransferase (MshA), dephosphorylation by the MSH phosphatase MshA2 (not yet identified), deacetylation by MshB to produce GlcN-Ins, linkage to Cys by the MSH ligase MshC, and acetylation by MSH synthase (MshD), yielding MSH. Studies of MSH mutants have shown that the MSH glycosyltransferase MshA and the MSH ligase MshC are required for MSH production, whereas mutants in the MSH deacetylase MshB and the acetyltransferase (MSH synthase) MshD produce some MSH and/or a closely related thiol. Current evidence indicates that MSH biosynthesis is controlled by transcriptional regulation mediated by σB and σR in Streptomyces coelicolor. Identified enzymes of MSH metabolism include mycothione reductase (disulfide reductase; Mtr), the S-nitrosomycothiol reductase MscR, the MSH S-conjugate amidase Mca, and an MSH-dependent maleylpyruvate isomerase. Mca cleaves MSH S-conjugates to generate mercapturic acids (AcCySR), excreted from the cell, and GlcN-Ins, used for resynthesis of MSH. The phenotypes of MSH-deficient mutants indicate the occurrence of one or more MSH-dependent S-transferases, peroxidases, and mycoredoxins, which are important targets for future studies. Current evidence suggests that several MSH biosynthetic and metabolic enzymes are potential targets for drugs against tuberculosis. The functions of MSH in antibiotic-producing streptomycetes and in bioremediation are areas for future study.

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“…Building on this model, Ferguson and co-workers used semi-quantitative complementation assays in conjunction with homology modeling to study the metal-binding and catalytic residues of rat PIG-L (12). From this data, they proposed a role for His-49 and Asp-52 of the AHPDD motif, along with the C-terminal His-157 of a HSNH motif, in metal binding.…”

Section: Discussionmentioning

confidence: 99%

Catalysis by N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase (PIG-L) from Entamoeba histolytica

Ashraf

Sreejith

Yadav

et al. 2013

Journal of Biological Chemistry

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Background: E. histolytica PIG-L is active even in absence of metal, unlike other homologs. Metal stimulation of activity alters V max , not K m . Metal does not alter optimum pH of catalysis. What explains these differences? Results: Conserved Asp-46 and His-140 participate in a general acid-base pair mechanism, unusual for de-N-acetylases. Conclusion: PIG-L of amoeba is significantly different from mammalian PIG-L. Significance: We have identified a probable drug target for selective delivery.

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The N-Acetyl-D-glucosaminylphosphatidylinositol De-N-acetylase of Glycosylphosphatidylinositol Biosynthesis Is a Zinc Metalloenzyme

Cited by 39 publications

References 38 publications

N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase from Entamoeba histolytica

N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase from Entamoeba histolytica

Biosynthesis and Functions of Mycothiol, the Unique Protective Thiol of Actinobacteria

Catalysis by N-Acetyl-d-glucosaminylphosphatidylinositol De-N-acetylase (PIG-L) from Entamoeba histolytica

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