Synthesis of myristoyl‐carba(dethia)‐coenzyme A and <i>S</i>‐(3‐oxohexadecyl)‐coenzyme A, two potent inhibitors of myristoyl‐CoA: protein <i>N</i>‐myristoyltransferase

Wagner, Aurel Popa; Rétey, János

doi:10.1111/j.1432-1033.1991.tb15756.x

Cited by 17 publications

(18 citation statements)

References 42 publications

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“…The traditional method for resolving isomers of CoA-containing compounds, ion exchange chromatography (1,2,14,34), is often lengthy, does not achieve base-line resolution, and thus may have hampered investigations into isomers of CoA (19). We report here an efficient HPLC methodology that achieves base line resolution of CoA and acyl-CoA isomers.…”

Section: Mass Spectrometric Structural Characterization Ofmentioning

confidence: 99%

“…The 2Ј-phosphate isomer of CoA, first named iso-CoA in the 1959 report of Moffatt and Khorana (1, 2), has traditionally been considered an undesirable synthetic by-product or has been simply ignored. Indeed, a review of the literature reveals that only eight of the more than 350 CoA-utilizing enzymes 2 have been examined for the ability to discriminate between the 2Ј-and 3Ј-phosphate isomers (1, 2, 10, 13, 14, 17-19, 34), and in two of these cases the enzymes have been shown to accept purified 2Ј-phosphate isomers as substrates or inhibitors (14,19). Moreover, the majority of the compounds examined in these eight cases are synthetic analogs of CoA, such as the dethia or seleno derivatives, and not isoCoA itself.…”

Section: Mass Spectrometric Structural Characterization Ofmentioning

confidence: 99%

“…In contrast, Thorpe et al (18) reported that substrate mixtures containing both dethia-isoCoA and dethia-CoA derivatives are capable of reducing enzyme-bound FAD. Likewise, Rossier (19) reported that choline acetyltransferase is inhibited by both seleno-CoA and selenoiso-CoA, and Wagner et al (14) have reported that N-myristoyltransferase is competitively inhibited by both carbadethiaCoA and carbadethia-iso-CoA, although in both cases the isoCoA analogs are less potent. In the cases of citrate synthase and carnitine palmitoyltransferase (17), it is difficult to draw a clear conclusion from the data because the experiments were performed with mixtures containing both the 2Ј-and 3Ј-isomers.…”

mentioning

confidence: 99%

“…These authors reported that HPLC 1 -purified samples of each isomer exhibited identical UV, HPLC-MS, and HPLC-MS/MS properties, and the identity of the 2Ј-phosphate isomer was convincingly demonstrated by comparing the one-dimensional NMR spectra to that of 2Ј,5Ј-ADP. In related work, Retey and co-workers have synthesized several dethia-CoA analogs (10 -15), in which the sulfur of CoA is replaced by a methylene, as mixtures of the 2Ј-and 3Ј-phosphate isomers; two of these analogs were purified to homogeneity and characterized by one-dimensional 1 H-NMR, 31 P-NMR, and, in one case, fast atom bombardment MS (13,14). Dethia-CoA analogs were also synthesized and characterized by Stewart and co-workers (16 -18); unfortunately, they were unable to separate the 2Ј-and 3Ј-phosphate isomers, and the NMR data could only be obtained on isomeric mixtures (17).…”

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Iso-coenzyme A

Burns

Gelbaum

Sullards

et al. 2005

Journal of Biological Chemistry

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Iso-coenzyme A is an isomer of coenzyme A in which the monophosphate is attached to the 2-carbon of the ribose ring. Although iso-CoA was first reported in 1959 (Moffatt, J. G., and Khorana, H. G. (1959) J. Am. Chem. Soc. 81, 1265-1265) to be a by-product of the chemical synthesis of CoA, relatively little attention has been focused on iso-CoA or on acyl-iso-CoA compounds in the literature. We now report structural characterizations of iso-CoA, acetyl-iso-CoA, acetoacetyl-iso-CoA, and ␤-hydroxybutyryl-iso-CoA using mass spectrometry (MS), tandem MS, and homonuclear and heteronuclear NMR analyses. Although the 2-phosphate isomer of malonyl-CoA was recently identified in commercial samples, previous characterizations of iso-CoA itself have been based on chromatographic analyses, which ultimately rest on comparisons with the degradation products of CoA and NADPH or have been based on assumptions regarding enzyme specificity. We describe a high performance liquid chromatography methodology for separating the isomers of several CoA-containing compounds. We also report here the first examples of isoCoA-containing compounds acting as substrates in enzymatic acyl transfer reactions. Finally, we describe a simple synthesis of iso-CoA from CoA, which utilizes ␤-cyclodextrin to produce iso-CoA with high regioselectivity, and we demonstrate a plausible mechanism that accounts for the existence of iso-CoA isomers in commercial preparations of CoA-containing compounds. We anticipate that these results will provide methodology and impetus for investigating iso-CoA compounds as potential pseudo-substrates or inhibitors of the >350 known CoA-utilizing enzymes.Iso-coenzyme A is an isomer of coenzyme A in which the monophosphate is attached to the 2Ј-carbon of the ribose ring. Iso-CoA was first reported in 1959 by Moffatt and Khorana (1, 2) to be a by-product of the chemical synthesis of CoA. The final step of this synthesis entailed acid-catalyzed hydrolysis of cyclic coenzyme A (Scheme 1) to produce a 50:50 mixture of two products that were separated using epicholorohydrin triethanolamine (ECTEOLA) cellulose ion exchange chromatography (1, 2). Direct structural analysis was not readily available in 1959; consequently, the authors deduced the structure of isoCoA by establishing that, in contrast to CoA, iso-CoA was not a substrate for the enzyme phosphotransacetylase (1, 2). Moffatt and Khorana also used paper chromatography of phosphodiesterase hydrolysates to show that enzymatic hydrolysis of isoCoA produces adenosine-2Ј,5Ј-diphosphate, exclusively (1, 2). Since the original work of Moffatt and Khorana was done, four additional chemical syntheses of CoA have been reported (3-8), all of which produce cyclic CoA as the penultimate product; cyclic CoA is then hydrolyzed either chemically to produce a mixture of CoA and iso-CoA (1, 2, 4 -6, 8) or enzymatically with ribonuclease T2 to produce CoA regioselectively (3-7).Recently, Minkler et al. (9) observed the 2Ј-phosphate isomer of malonyl-CoA in commercial samples. These authors ...

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Section: Mass Spectrometric Structural Characterization Ofmentioning

confidence: 99%

Section: Mass Spectrometric Structural Characterization Ofmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Iso-coenzyme A

Burns

Gelbaum

Sullards

et al. 2005

Journal of Biological Chemistry

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“…The pioneering work of Gordon and colleagues led to the design and synthesis of antifungal peptidomimetic and dipeptide analog inhibitors for C. albicans NMT that are selective versus human NMT while demonstrating low nanomolar potencies toward the enzyme (Devadas et al, 1995Nagarajan et al, 1997). Other recently described inhibitors include amino acid derivatives (Boutin et al, 1993;Furuishi et al, 1997;Tabuchi et al, 1997;Cordo et al, 1999) and nonhydrolyzable myristoyl-CoA analogs Paige et al, 1989;Wagner and Retey, 1991). However, the development of these compounds may be limited by their poor pharmacokinetic properties.…”

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Cyclohexyl-octahydro-pyrrolo[1,2-a]pyrazine-Based Inhibitors of HumanN-Myristoyltransferase-1

French¹,

Zhuang²,

Schrecengost³

et al. 2004

J Pharmacol Exp Ther

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N-Myristoyltransferase (NMT) is an emerging therapeutic target that catalyzes the attachment of myristate to the N terminus of an acceptor protein. We have developed a medium-throughput assay for screening potential small molecule inhibitors of human NMT-1 consisting of recombinant enzyme, biotinylated peptide substrate, and [3 H]myristoyl-CoA. Approximately 16,000 diverse compounds have been evaluated, and significant inhibition of NMT was found with 0.8% of the compounds. From these hits, we have identified the cyclohexyl-octahydropyrrolo[1,2-a]pyrazine (COPP) chemotype as inhibitory toward human NMT-1. Thirty-two compounds containing this substructure inhibited NMT-1, with IC 50 values ranging from 6 M to millimolar concentrations, and a quantitative structure-activity relationship equation (r 2 ϭ 0.72) was derived for the series. The most potent inhibitor (24, containing 9-ethyl-9H-carbazole) demonstrated competitive inhibition for the peptide-binding site of NMT-1 and noncompetitive inhibition for the myristoylCoA site. Computational docking studies using the crystal structure of the highly homologous yeast NMT confirmed that 24 binds with excellent complementarity to the peptide-binding site of the enzyme. To evaluate the ability of 24 to inhibit NMT activity in intact cells, monkey CV-1 cells expressing an Nmyristoylated green fluorescent protein (GFP) fusion protein were treated with a known NMT inhibitor or with 24. Each compound caused the redistribution of GFP from the plasma membrane to the cytosol. Furthermore, 24 inhibits cancer cell proliferation at doses similar to those that inhibit protein myristoylation. Overall, these studies establish an efficient assay for screening for inhibitors of human NMT and identify a novel family of inhibitors that compete at the peptide-binding site and have activity in intact cells.The term protein myristoylation refers to the covalent attachment of myristic acid to specific proteins. This process has been shown to target proteins to specific subcellular membranes (Boutin, 1997), stabilize protein conformation (Chow et al., 1987;Zheng et al., 1993;Resh, 1996), and facilitate additional lipidations (Robbins et al., 1995;Yurchak and Sefton, 1995;Song et al., 1997;van't Hof and Resh, 1999). Although the reaction mainly occurs as a cotranslational event, it has been shown to occur after cleavage of some substrates (Zha et al., 2000). N-Myristoyltransferase (NMT; E.C. 2.3.1.97) catalyzes this addition onto the N-terminal glycine residue of specific proteins. The reaction requires only myristoyl-CoA and a protein containing a suitable peptide sequence and occurs through an ordered Bi Bi mechanism Rocque et al., 1993;Bhatnagar et al., 1998). The list of myristoylated proteins is diverse, including viral proteins, G␣ proteins, tyrosine kinases of the Src family, and nitric oxide synthase. Gene replacement studies indicate that NMT is essential for viability of Cryptococcus neoformans (Lodge et al., 1994). In addition, mutation or deletion of NMT results in recess...

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N-Ethyl-5-phenylisoxazolium-3′-sulfonate

Pottorf¹

2001

Encyclopedia of Reagents for Organic Synthesis

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Synthesis of myristoyl‐carba(dethia)‐coenzyme A and S‐(3‐oxohexadecyl)‐coenzyme A, two potent inhibitors of myristoyl‐CoA: protein N‐myristoyltransferase

Cited by 17 publications

References 42 publications

Iso-coenzyme A

Iso-coenzyme A

Cyclohexyl-octahydro-pyrrolo[1,2-a]pyrazine-Based Inhibitors of HumanN-Myristoyltransferase-1

N-Ethyl-5-phenylisoxazolium-3′-sulfonate

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