Backbone Cyclic Peptide, Which Mimics the Nuclear Localization Signal of Human Immunodeficiency Virus Type 1 Matrix Protein, Inhibits Nuclear Import and Virus Production in Nondividing Cells

Friedler, Assaf; Zakai, Nehama; Karni, Orit; Broder, Yehoshua C.; Baraz, Lea; Kotler, Moshe; Loyter, A.; Gilon, Chaim

doi:10.1021/bi972878h

Cited by 59 publications

(71 citation statements)

References 33 publications

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“…The coupling of all the amino acids following this arginine were repeated twice. Succinic and glutaric acids were coupled as their anhydrides (10 eq) with 10 eq diisopropylethylamine and 1 eq dimethylaminopyridine in NMP for 3 h. Couplings of adipic and pimelic acid were performed with 10 eq acid and 10 eq DIC, which were preactivated in DMF for 30 min and then added to the resin with 1 eq dimethylaminopyridine and shaken for 3 h. HF cleavage and work up were performed as described (13). The crude peptides were analyzed by time of flight-mass spectroscopy and had the expected molecular weights.…”

Section: Methodsmentioning

confidence: 99%

“…13. Ac-GRKKRRQRRRAHQN-NH 2 was used for the linear Tat-ARM; this sequence has been shown by Siomi et al (26) to act as an NLS.…”

Section: Synthesis Of Linear Peptides-linearmentioning

confidence: 99%

“…Quantitative Analysis of Nuclear Import in an in Vitro SystemNuclear import was quantitatively determined by the ELISA-based assay system using biotinylated BSA peptide conjugates as transport substrates, essentially as described before (13,27). Briefly, a suspension of Colo-205 cells was used, and biotinylated BSA-SV40 T-antigen NLS, biotinylated BSA-Tat NLS, and biotinylated BSA-ARM mimetic BC peptides conjugates were used as transport substrates.…”

Section: Synthesis Of Linear Peptides-linearmentioning

confidence: 99%

“…We have demonstrated its utility with the parent proteins bovine pancreatic trypsin inhibitor (1,11,12) and HIV-1 matrix protein nuclear localization signal (NLS) (13).…”

mentioning

confidence: 99%

“…However, BCvir was not able to mediate nuclear import of the appropriate conjugate. It inhibited the function of the corresponding receptor but was not able to activate it, showing an antagonistic activity (13,24). BCvir is not an optimal NLS mimetic because it does not display the properties of an active NLS (agonistic activity).…”

mentioning

confidence: 99%

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Development of a Functional Backbone Cyclic Mimetic of the HIV-1 Tat Arginine-rich Motif

Friedler¹,

Friedler²,

Luedtke³

et al. 2000

Journal of Biological Chemistry

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We have used the backbone cyclic proteinomimetics approach to develop peptides that functionally mimic the arginine-rich motif (ARM) of the HIV-1 Tat protein. This consensus sequence serves both as a nuclear localization signal (NLS) and as an RNA binding domain. Based on the NMR structure of Tat, we have designed and synthesized a backbone cyclic ARM mimetic peptide library. The peptides were screened for their ability to mediate nuclear import of the corresponding BSA conjugates in permeabilized cells. One peptide, designated "Tat11," displayed active NLS properties. Nuclear import of Tat11-BSA was found to proceed by the same distinct pathway used by the Tat-NLS and not by the common importin ␣ pathway, which is used by the SV40-NLS. Most of the Tat-derived backbone cyclic peptides display selective inhibitory activity as demonstrated by the inhibition of the nuclear import mediated by the Tat-NLS and not by the SV40-NLS. The Tat-ARM-derived peptides, including Tat-11, also inhibited binding of the HIV-1 Rev-ARM to its corresponding RNA element (Rev response element) with inhibition constants of 5 nM. Here we have shown for the first time (a) a functional mimetic of a protein sequence, which activates a nuclear import receptor and (b) a mimetic of a protein sequence with a dual functionality. Tat11 is a lead compound which can potentially inhibit the HIV-1 life cycle by a dual mechanism: inhibition of nuclear import and of RNA binding.Proteinomimetics are small molecules that can mimic the structure and/or function of active sites within proteins (1). They are useful for detailed study of protein folding, structure, and function. The major potential application of proteinomimetics is, however, therapeutic; such molecules can be used to block protein-protein and/or protein-nucleic acid interactions, thus interfering with undesired biological processes (2-4). Being relatively small, proteinomimetics often solve acute problems associated with the use of proteins as drugs, such as antigenicity, low bioavailability, high cost, and rapid enzymatic degradation.Two properties of the parent protein must be retained when designing proteinomimetics: (i) the bioactive conformation of the desired active site and (ii) a certain degree of conformational flexibility to allow induced fit. Linear peptides are not optimal candidates to mimic proteins, because they equilibrate between multiple conformations and thus adaptation of the bioactive conformation is at an entropic cost. Introduction of conformational constraints into the peptide is thus needed to generate a proteinomimetic. Being relatively small and conformationally constrained, cyclic peptides are excellent candidates to serve as proteinomimetics. Backbone cyclization of peptides is the method of cyclization developed and used in our lab. It results in peptides with improved selectivity, enhanced metabolic stability, and high bioavailability (5-9). To select the most active backbone cyclic (BC) 1 peptide based on a given sequence, we have developed the "cycloscan" ...

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Section: Methodsmentioning

confidence: 99%

“…13. Ac-GRKKRRQRRRAHQN-NH 2 was used for the linear Tat-ARM; this sequence has been shown by Siomi et al (26) to act as an NLS.…”

Section: Synthesis Of Linear Peptides-linearmentioning

confidence: 99%

Section: Synthesis Of Linear Peptides-linearmentioning

confidence: 99%

“…We have demonstrated its utility with the parent proteins bovine pancreatic trypsin inhibitor (1,11,12) and HIV-1 matrix protein nuclear localization signal (NLS) (13).…”

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Development of a Functional Backbone Cyclic Mimetic of the HIV-1 Tat Arginine-rich Motif

Friedler¹,

Friedler²,

Luedtke³

et al. 2000

Journal of Biological Chemistry

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Resistance to Inhibitors of HIV‐1 Integrase

Hombrouck¹,

Clayton²,

Voet

et al. 2011

HIV‐1 Integrase

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Structure‐activity relationship and metabolic stability studies of backbone cyclization and N‐methylation of melanocortin peptides

et al. 2008

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Backbone cyclization (BC) and N-methylation have been shown to enhance the activity and/or selectivity of biologically active peptides and improve metabolic stability and intestinal permeability. In this study, we describe the synthesis, structure-activity relationship (SAR) and intestinal metabolic stability of a backbone cyclic peptide library, BL3020, based on the linear alpha-Melanocyte stimulating hormone analog Phe-D-Phe-Arg-Trp-Gly. The drug lead, BL3020-1, selected from the BL3020 library (compound 1) has been shown to inhibit weight gain in mice following oral administration. Another member of the BL3020 library, BL3020-17, showed improved biological activity towards the mMC4R, in comparison to BL3020-1, although neither were selective for MC4R or MC5R. N-methylation, which restrains conformational freedom while increasing metabolic stability beyond that which is imparted by BC, was used to find analogs with increased selectivity. N-methylated backbone cyclic libraries were synthesized based on the BL3020 library. SAR studies showed that all the N-methylated backbone cyclic peptides demonstrated reduced biological activity and selectivity for all the analyzed receptors. N-methylation of active backbone cyclic peptides destabilized the active conformation or stabilized an inactive conformation, rendering the peptides biologically inactive. N-methylation of backbone cyclic peptides maintained stability to degradation by intestinal enzymes.

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Backbone Cyclic Peptide, Which Mimics the Nuclear Localization Signal of Human Immunodeficiency Virus Type 1 Matrix Protein, Inhibits Nuclear Import and Virus Production in Nondividing Cells

Cited by 59 publications

References 33 publications

Development of a Functional Backbone Cyclic Mimetic of the HIV-1 Tat Arginine-rich Motif

Development of a Functional Backbone Cyclic Mimetic of the HIV-1 Tat Arginine-rich Motif

Resistance to Inhibitors of HIV‐1 Integrase

Structure‐activity relationship and metabolic stability studies of backbone cyclization and N‐methylation of melanocortin peptides

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