Designing photoaffinity labeling reagents for chromatin studies

Buchardt, Ole; Ehrbar, Ulrich; Larsen, Charles; Moeller, J.; Nielsen, Peter E.; Thomsen, T.; Waetjen, F.; Hansen, John Bondo

doi:10.1021/jo00196a004

Cited by 16 publications

(7 citation statements)

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“…The in vitro cytotoxicities of the compounds were determined as described previously29 against the murine leukemia P388 and Chinese hamster ovary derived AA8 and UV4 cell lines and are given in Table I. The UV4 subline of AA830,31 lacks the ability to perform the incision step of the excision repair pathway of DNA damage repair, and these cells are consequently from 2-10-fold more sensitive than AA8 to compounds, which form bulky DNA monoadducts, and from 8-200-fold more sensitive than AA8 to DNA interstrand cross-linking agents.32 The ratio of the ICijo values of a drug in these two cell lines (hypersensitivity factor, HF) can thus provide information (29) 0 ICso values determined against either P388 or AA8 cells, as described in ref 22 and outlined in the text, and the standard error of the mean. 6HF = hypersensitivity factor = ICM (AASI/ICjq (UV4), as outlined in the text.…”

Section: Biological Activitymentioning

confidence: 99%

“…Methyl 3-[(4-Aminophenyl)thio]propanoate (22). A suspension of methyl 3-[(4-nitrophenyl)thio]propanoate (21)18 (23.3 g, 0.10 mmol) and SnCl2-2H20 (109 g, 0.48 mol) in EtOAc (500 mL) was heated under reflux for 4 h under N2.…”

Section: S-(benzyloxycarbonyl)-4-[ethyl(2-hydroxyethyl)-mentioning

confidence: 99%

“…The cooled mixture was basified with ammonia and filtered, and the precipitate was washed repeatedly with EtOAc. The combined filtrates were washed with water and worked up to give methyl 3-[(4-aminophenyl)thio]propanoate (22) as an orange oil (17.1 g, 84%): *H NMR (CDC13; 60 MHz) b 2.53 (t, J = 4.3 Hz, 2 H, CH2CO), 2.96 (t, J = 5.8 Hz, 2 H, SCH2), 3.62 (s, 5 H, OMe, NH2), 6.55 (d, J = 8.8 Hz, 2 H, H-3,5), 7.22 (d, J = 9.1 Hz,2 H,6). The compound was used immediately without further purification.…”

Section: S-(benzyloxycarbonyl)-4-[ethyl(2-hydroxyethyl)-mentioning

confidence: 99%

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Synthesis and evaluation of DNA-targeted spatially separated bis(aniline mustards) as potential alkylating agents with enhanced DNA cross-linking capability

Gourdie¹,

Prakash²,

Wakelin³

et al. 1991

J. Med. Chem.

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DNA-targeted separated bis-mustards were synthesized by attaching aniline mono-mustards at the 4- and 9-positions of the DNA-intercalating ligand 9-aminoacridine-4-carboxamide, with the intention of improving the low cross-link to monoadduct ratio found with most alkylating agents. The geometry of these compounds requires that, when the acridine binds to DNA by intercalation, one alkylating moiety is delivered to each DNA groove. Gel electrophoretic studies show that only one arm of these compounds (probably that attached to the 9-position) alkylates DNA, such alkylation occurring specifically in the major groove at the N7 of guanines. Cell-line studies confirm that the mode of cytotoxicity of these compounds (unlike that of untargeted aniline bis-mustards of comparable reactivity) is due to bulky DNA monoadduct formation. It is concluded that more information is required about the exact orientation of the initial monoadducts before ligands with specific DNA cross-linking ability can be designed.

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Section: Biological Activitymentioning

confidence: 99%

Section: S-(benzyloxycarbonyl)-4-[ethyl(2-hydroxyethyl)-mentioning

confidence: 99%

Section: S-(benzyloxycarbonyl)-4-[ethyl(2-hydroxyethyl)-mentioning

confidence: 99%

See 1 more Smart Citation

Synthesis and evaluation of DNA-targeted spatially separated bis(aniline mustards) as potential alkylating agents with enhanced DNA cross-linking capability

Gourdie¹,

Prakash²,

Wakelin³

et al. 1991

J. Med. Chem.

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“…As an extension of our efforts to design photochemical probes for the study of protein-DNA interactions [17][18][19][20][21][22] we now report that azidoacridine-DNA photoadducts can be cleaved with piperidine and we show that 9-amino-6-azido-2-methoxyacridines can be used as DNA-photocleaving and DNA-photofootprinting reagents.…”

Section: ] Andmentioning

confidence: 85%

Photocleavage of DNA and photofootprinting of E.coli RNA polymerase bound to promoter DNA by azido-9-acridinylamines

Jeppesen¹,

Buchardt

Henriksen

et al. 1988

Nucleic Acids Research

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The long-wavelength ultraviolet (lambda approximately 420 nm) radiation induced reaction between 6-azido-2-methoxy-9-acridinylamines and supercoiled plasmid DNA results in single strand scissions and formation of covalent adducts (ratio approximately 1:10). By treating azidoacridine-photomodified DNA with piperidine at 90 degrees C, additional strand scissions are observed in a complex sequence dependent manner with an overall preference for T greater than or equal to G greater than C much greater than A. The resulting DNA fragments migrate as 5'-phosphates in polyacrylamide gels. Photofootprinting of the binding site of RNA-polymerase on promoter DNA is demonstrated with an azido-9-acridinylamino-octamethylene-9-aminoacridine. Similar experiments using 9-amino-6-azido-2-methoxyacridine indicate that this reagent recognizes changes in the DNA conformation induced by RNA polymerase binding, in relation to open complex formation.

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“…strong reversible binding to DNA by intercalation, and a photoprobe, e.g., an aryl azide group, which reacts irreversibly in situ on irradiation with long-wavelength u.v. light (Nielsen, 1981(Nielsen, , 1982Nielsen et al, 1983; Buchardt et al, 1984).…”

mentioning

confidence: 99%

Photochemical cross-linking of protein and DNA in chromatin. Synthesis and application of a photosensitive cleavable derivative of 9-aminoacridine with two photoprobes connected through a disulphide-containing linker

Nielsen¹,

Hansen²,

Buchardt³

1984

Biochemical Journal

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A novel cleavable photo-cross-linking reagent, N-(2-methoxy-6-azidoacridin-9-yl)-N'-(4-azidobenzoyl)cystamine, for analysis of protein-nucleic acid interactions, has been synthesized. The reagent contains two photosensitive groups that can be activated sequentially. The azidoacridinyl moiety is sensitive to u.v. and visible light (lambda less than or equal to 450 nm), whereas the azidobenzoyl part needs higher-energy light (lambda less than or equal to 350 nm). Furthermore, the disulphide bridge connecting the two photoactive groups can be cleaved by reduction with mercaptans. The reagent is shown to induce cleavable cross-links between all five major histones and DNA in chromatin from Ehrlich ascites cells on activation with long-wavelength u.v. light (lambda greater than 300 nm) at an efficiency of approximately 3% of the added reagent.

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Designing photoaffinity labeling reagents for chromatin studies

Cited by 16 publications

References 0 publications

Synthesis and evaluation of DNA-targeted spatially separated bis(aniline mustards) as potential alkylating agents with enhanced DNA cross-linking capability

Synthesis and evaluation of DNA-targeted spatially separated bis(aniline mustards) as potential alkylating agents with enhanced DNA cross-linking capability

Photocleavage of DNA and photofootprinting of E.coli RNA polymerase bound to promoter DNA by azido-9-acridinylamines

Photochemical cross-linking of protein and DNA in chromatin. Synthesis and application of a photosensitive cleavable derivative of 9-aminoacridine with two photoprobes connected through a disulphide-containing linker

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