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Monoclonal antibodies against pools of mono- and polyacetylated peptides selectively recognize acetylated lysines within the context of the original antigen (85 visite) (PDF privato 110 visite)

Sandomenico A, Focà A, Sanguigno L, Caporale A, Focà G, Pignalosa A, Corvino G, Caragnano A, Beltrami AP, Antoniali G, Tell G, Leonardi A, Ruvo M

Medicine Research And Experimental (ISSN: 1942-0862, 1942-0870e), 2016 Aug; 8: 1575-1589.

Tipo di articolo: Journal Article,

Impact factor: 4.881

Impact factor a 5 anni: 5.07

Parole chiave: Acetylation; Anti-Acetyl-Peptide Monoclonal Antibodies; Peptide Libraries; Ape1; Ref-1; Acetylated-Ape1; Human Apurinic, Apyrimidinic Endonuclease-1; N-Terminal Domain; Protein; Ape1; Ape1, Ref-1; Binding; Cells; Inhibition; Metabolism; Interacts,

Url: Non disponibile.

Post-translational modifications (PTMs) strongly influence the structure
and function of proteins. Lysine side chain acetylation is one of the
most widespread PTMs, and it plays a major role in several physiological
and pathological mechanisms. Protein acetylation may be detected by
mass spectrometry (MS), but the use of monoclonal antibodies (mAbs) is a
useful and cheaper option. Here, we explored the feasibility of
generating mAbs against single or multiple
acetylations within the context of a specific sequence. As a model, we
used the unstructured N-terminal domain of APE1, which is acetylated on
Lys27, Lys31, Lys32 and Lys35. As immunogen, we used a peptide mixture
containing all combinations of single or multi-acetylated variants
encompassing the 24-39 protein region. Targeted screening of the
resulting clones yielded mAbs that bind with high affinity to only the
acetylated APE1 peptides and the acetylated protein. No binding was seen
with the non-acetylated variant or unrelated acetylated peptides and
proteins, suggesting a high specificity for the APE1 acetylated
molecules. MAbs could not finely discriminate between the differently
acetylated variants; however, they specifically bound the acetylated
protein in mammalian cell extracts and in intact cells and tissue slices
from both breast cancers and from a patient affected by idiopathic
dilated cardiomyopathy. The data suggest that our approach is a rapid
and cost-effective method to generate mAbs against specific proteins
modified by multiple acetylations or other PTMs.
*** IBB - CNR ***

Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche (IBB-CNR), Napoli, Italy;

Centro Interuniversitario di Ricerca sui Peptidi Bioattivi (CIRPeB), Napoli, Italy;

Bioker Multimedica, Napoli, Italy;
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E Data iniziale: Data finale: [X]   E Sede:  E    Affiliazione IBB  
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Sandomenico A, Caporale A, Doti N, Cross S, Cruciani G, Chambery A, De Falco S, Ruvo M
* Synthetic Peptide Libraries. From random mixtures to in vivo testing (9 visite)
Curr Med Chem (ISSN: 1875-533xelectronic, 0929-8673linking), 2018 Jul 15; N/D: N/D-N/D.
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Magrì A, Tabbì G, Giuffrida A, Pappalardo  G, Satriano C, Naletova I, Nicoletti VG, Attanasio F
* Influence of the N-terminus acetylation of Semax, a synthetic analog of ACTH(4-10), on copper(II) and zinc(II) coordination and biological properties (86 visite)
J Chem Res (ISSN: 1873-3344electronic, 0162-0134linking, 0162-0134print), 2016 Aug 27; 164: 59-69.
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Pellerito O, Prinzivalli C, Foresti E, Sabatino P, Abbate M, Casella G, Fiore T, Scopelliti M, Pellerito C, Giuliano M, Grasso G, Pellerito L
* Synthesis, chemical characterization and biological activity of new histone acetylation/deacetylation specific inhibitors: A novel and potential approach to cancer therapy (128 visite)
J Chem Res (ISSN: 0162-0134, 1873-3344, 0162-0134print), 2013 Sep; 125: 16-25.
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Poletto M, Vascotto C, Scognamiglio PL, Lirussi L, Marasco D, Tell G
* Role Of The Unstructured N-Terminal Domain Of The Human Apurinic/Apyrimidinic Endonuclease 1 (hape1) In The Modulation Of Its Interaction With Nucleic Acids And Nucleophosmin (npm1) (116 visite)
Biochem J (ISSN: 0264-6021, 1470-8728electronic, 0264-6021linking), 2013; 452(3): 545-557.
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Lirussi L, Antoniali G, Vascotto C, D'Ambrosio C, Poletto M, Romanello M, Marasco D, Leone M, Quadrifoglio F, Bhakat KK, Scaloni A, Tell G
* Nucleolar accumulation of APE1 depends on charged lysine residues that undergo acetylation upon genotoxic stress and modulate its BER activity in cells (92 visite)
Molecular Biology Of The Cell (ISSN: 1059-1524), 2012 Oct 15; 23(20): 4079-4096.
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Poletto M, Di Loreto C, Marasco D, Poletto E, Puglisi F, Damante G, Tell G
* Acetylation on critical lysine residues of Apurinic/apyrimidinic endonuclease 1 (APE1) in triple negative breast cancers (96 visite)
Biochemical And Biophysical Research Communications (ISSN: 0006-291x), 2012 Jul 20; 424(1): 34-39.
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Doti N, Scognamiglio PL, Madonna S, Scarponi C, Ruvo M, Perretta G, Albanesi C, Marasco D
* New mimetic peptides of the kinase-inhibitory region (KIR) of SOCS1 through focused peptide libraries (128 visite)
Biochemical Journal, 2012 Apr 1; 443: 231-240.
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Canettieri G, Di Marcotullio L, Greco A, Coni S, Antonucci L, Infante P, Pietrosanti L, De Smaele E, Ferretti E, Miele E, Pelloni M, De Simone G, Pedone E, Gallinari P, Giorgi A, Steinkuhler C, Vitagliano L, Pedone C, Schinin ME, Screpanti I, Gulino A
* Histone deacetylase and Cullin3-REN(KCTD11) ubiquitin ligase interplay regulates Hedgehog signalling through Gli acetylation (119 visite)
Nature Cell Biol Nature Cell Biology (ISSN: 1476-4679, 1465-7392), 2010 Feb; 12(2): 132-142.
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Marasco D, Perretta G, Sabatella M, Ruvo M
* Past and Future Perspectives of Synthetic Peptide Libraries (102 visite)
Curr Protein Pept Sci (ISSN: 1389-2037), 2008 Oct; 9(5): 447-467.
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9 Records (9 escludendo Abstract e Conferenze).
Impact factor totale: 50.437 (50.437 escludendo Abstract e Conferenze).
Impact factor a 5 anni totale: 52.576 (52.576 escludendo Abstract e Conferenze).

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