Synthesis and characterization of the Nω , Nω -dimethylarginine rich C-terminal region of human nucleolin (CROSBI ID 492983)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | međunarodna recenzija
Podaci o odgovornosti
Zahariev, Sotir ; Guarnaccia, Corrado ; Zanuttin, Francesco ; Pintar, Alessandro ; Maravić, Gordana ; Pongor, Sandor
engleski
Synthesis and characterization of the Nω , Nω -dimethylarginine rich C-terminal region of human nucleolin
Both versions of C-terminal region of human nucleolin ChNu (646-706), containing Arg (I), respectively Nω , Nω -dimethylarginine (II), from the family of RGG-rich proteins, were successfully synthesized by an automated Fmoc methodology for SPPS using dimethoxybenzyl as a reversible amide bond protecting group. The cleavage/deprotection conditions were optimized and ChNu's were purified to homogeneity >95% by a combination of IE- and RP-HPLC. The purity/identity of peptides were confirmed by ESI-MS, AAA, SDS-PAGE, RP HPLC in two different systems and by IE HPLC. Arg-dimethylation increases hydrophobicity, as determined by RP-HPLC, and decreases basicity (IE-HPLC). In double filter binding assays (I) and (II) show very similar binding potency to single-stranded DNA (ssDNA) with values comparable to that of recombinant fusion ChNu (Kd ~ 2X10-7 M). The nucleic acid binding affinity is substantially increased in comparison to our published result on short peptides and correlates with the higher number of RGG, respectively aDma-GG-repeats. In a ssDNA-agarose affinity chromatography experiment Arg-dimethylation decreased the electrostatic interaction with ssDNA. Both (I) and (II) show very similar binding potency to G4 DNA and do not exhibit helicase activity in a M13 phage assay. (I) is efficiently, but not exhaustively methylated in vitro by recombinant protein arginine N-methyltransferase type I. 1H NMR spectra of (I) and (II) are characterized by the lack of dispersion for NH and Hα chemical shifts suggesting the absence of globular structure.
nucleolin; peptide synthesis; dimethylation; arginine
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Podaci o prilogu
336-337.
2002.
objavljeno
Podaci o matičnoj publikaciji
Benedetti, Ettore ; Pedone, Carlo
Napulj: Edizioni Ziino
88-900948-1-8
Podaci o skupu
27th European Peptide Symposium
poster
01.01.2002-01.01.2002
Sorrento, Italija