In Vitro Characterization of Peptides Interfering with the HIV-1 Nucleocapsid Protein (NCp7) Functions. Pp. 299-306.
Daniela Lener, Giovanni Antico, Alssandra André, Anna Aulicino, Enrico Bucci,
Jean-Luc Darlix and Raffaele A. Calogero
[Abstract]
FMOC- and NSC-Groups as a Base Labile N(alpha)-Amino Protection: A Comparative
Study in the Automated SPPS. Pp. 307-312.
Aydar N. Sabirov, Young-Deug Kim, Hack- Joo Kim and Vladimir V. Samukov
[Abstract]
Identification of the Isoforms and Subisoforms of Rabbit Liver Metallothionein
Using Electrospray Mass Spectrometry. Pp. 313-320.
J. C. Yves Le Blanc, Anthony Presta, John Veinot, Dean Gibson, K. W. Michael Siu
and Martin J. Stillman
[Abstract]
Mutant of HIV-1 Protease with New Specific properties. Pp. 321-328.
Dergousova, N.I., Leonova, Yu. F., Zinchenko, A.A., Rumsh, L.D. and
Andreeva, N.S.
[Abstract]
Crystal Structure of Myotoxin-II: A Myotoxic Phospholipase A2 - Homologue from Bothrops Moojeni Venom. Pp. 329-334.
W.F. de Azevedo Jr., R.f Ward, F.R. Lombardi, J.R. Gigho, A.M. Soares, M.R.M. Fontes and R.K. Arni
[Abstract]
Introduction of Trypsin Specificity into Chymotrypsin Inhibitor II . Pp. 335-342.
Zulfiqar Hasan and Robin J. Leatherbarrow
[Abstract]
Crystallisation Reports
Crystallization and Preliminary Crystallographic Analysis of Scilla campanalata Lectin
Complexed with alpha-D-Mannose. Pp. 343-348.
Lisa M. Wright, Stephen D. Wood, Colin D. Reynolds, Pierre J. Rizkallah and Anthony K. Allen
[Abstract]
Crystallization and X-Ray Diffraction Data Analysis of Oxyhemoglobin-I from the
Catfish Liposarcus anisitsi (PISCES). Pp. 349-354.
Smarra, A.L.S., Arni, R.K., De Azevedo, Jr., W.F. Colombo, M.F., and
Bonilla-Rodriguez, G. 0.
[Abstract]
[Back to top] Interaction of a Partial Calmodulin-Binding Domain of Caldesmon with Calmodulin. Mingjie Zhang and Hans J. Vogel.
Caldesmon (CaD) is associated with the thin filaments of smooth muscle tissues. Limited proteolysis studies of CaD reveal that the sequence W659 EKGNVF is involved in binding the calcium-regulatory protein calmodulin (CaM). Using transferred nuclear Overhauser enhancement NMR measurements, we have found that the 8 and 11 residue synthetic peptides KS(MWEKGNVF)S bind with an alpha-helical structure to CaM. Proton NMR titration studies show that a 2:1 peptide to CaM complex is formed, suggesting that the two domains of the dumbbell-shaped CaMcan bind one equivalent of peptide each with sirnilar binding constant.
[Back to top] In Vitro Characterization of Peptides Interfering with the HIV-1 Nucleocapsid Protein (NCp7) Functions. Daniela Lener, Giovanni Antico, Alssandra André, Anna Aulicino, Enrico Bucci,
Jean-Luc Darlix and Raffaele A. Calogero.
In HIV-1, the nucleocapsid protein (NCp7) plays essential roles in several steps of HIV-1 replication. Since NCp7 is required for virion formation and proviral DNA synthesis and is highly conserved, identification of compounds able to inhibit NCp7 activities and functions may lead to the discovery of new anti-HIV drugs. Here we present data showing that peptides derived from the first and second NCp7 zinc finger interfere, in vitro, with NCp7 functions during the early stage of HIV-1 reverse transcription.
[Back to top] FMOC- and NSC-Groups as a Base Labile N(alpha)-Amino Protection: A Comparative Study in the Automated SPPS. Aydar N. Sabirov, Young-Deug Kim, Hack- Joo Kim and Vladimir V. Samukov.
Base-labile 2-(4-nitrophenylsulfonyl)ethoxycarbonyl (Nsc) and 9-fluorenylmethoxycarbonyl (Fmoc) groups are compared as a temporary N(alpha)-amino protection in the automated SPPS of WN-alpha2(130-137), ACP(65-74) and ACTH(1-24) on MilliGen 9050+ PepSynthesizer(TM). The syntheses were performed on PEG-PS and Wang PS resins using standard BOP/DIEA protocol with some modifications. HPLC profiles of the crude peptides and isolated yields resulted from both Nsc and Fmoc assemblies were almost identical for the three test peptides.
[Back to top] Identification of the Isoforms and Subisoforms of Rabbit Liver Metallothionein Using Electrospray Mass Spectrometry. J. C. Yves Le Blanc, Anthony Presta, John Veinot, Dean Gibson, K. W. Michael Siu
and Martin J. Stillman.
A cartridge-based, chromatographic method is reported for the isolation of isoform 2a of rabbit liver zinc metallothionein that reduces the time of key steps by 50-fold and is scaleable from analytical (<5 ppb zinc) to preparative (100's ppm zinc) levels. Electrospray mass spectra are reported that unambiguously identify for the first time the presence of each of the 6 subisoforms of rabbit liver metallothionein. These data show that MT 2a elutes with minor fractions of 2b and 2c, whereas MT 1a elutes with major fractions of 2d and 2e.
[Back to top] Mutant of HIV-1 Protease with New Specific properties. Dergousova, N.I., Leonova, Yu. F., Zinchenko, A.A., Rumsh, L.D. and
Andreeva, N.S.
An extensive net of hydrogen bonds includuig the active carboxyls and Tyr residue locating in the so-called 'flap' in pepsin-like enzymes was planned to be restored in moleculle of HIV-1 protease. For this purpose, 5 mutations and 2 deletions were introduced simultaneously in the enzyme. Some of the residues subjected to mutagenesis are known to be changed duririg formation of drug resistant HIV-1 protease. The obtained mutant was fully active, but had an altered specificity and pH optimum.
[Back to top] Crystal Structure of Myotoxin-II: A Myotoxic Phospholipase A2 - Homologue from Bothrops Moojeni Venom. W.F. de Azevedo Jr., R.f Ward, F.R. Lombardi, J.R. Gigho, A.M. Soares, M.R.M. Fontes and R.K. Arni.
The crystal structure of Myotoxin-II (MjTX-II), a Lys49 PLA2-homologue from Bothrops moojeni venom has been determined and refined at 2.0 A to a crystallographic residual of 19.7% (Rfree=28.1%). MjTX-II is a dimer in the crystal, with the monomers in the asymmetric unit related by a two-fold symmetry axis running through the dimer interface. The dimers of MjTX-II and the Lys49 PLA2 from B. asper venom are similar, however the relative orientations of the monomers suggests a flexible dimer interface, which serves as a hinge between the two molecules.
[Back to top] Introduction of Trypsin Specificity into Chymotrypsin Inhibitor II. Zulfiqar Hasan and Robin J. Leatherbarrow.
A unique cysteine residue was introduced into the P1 position of chymotrypsin inhibitor II (CI2) by
oligonucleotide-directed mutagenesis (Met59 -> Cys). Chemical modification of CI2M59C with 2-bromoethylamine, 3-bromopropylamine and 2-mercaptoethylamine resulted in the introduction of lysine-sulfur analogues at the P1 position. Unlike the original CI2, all three modified inhibitors exhibited moderate but effective inhibition of trypsin. The results demonstrate that genetic and chemical modification can be successfully combined to tailor inhibitory specificity.
[Back to top] Crystallization and Preliminary Crystallographic Analysis of Scilla campanalata Lectin Complexed with alpha-D-Mannose. Lisa M. Wright, Stephen D. Wood, Colin D. Reynolds, Pierre J. Rizkallah and Anthony K. Allen.
Recently many new lectins have been isolated from monocotyledonous plants which have interesting glycan-binding behaviour. In this paper we present the crystallization and preliminary X-ray analysis of a lectin, isolated from bluebell (Scilla campanulata) bulbs, complexed with alpha-D-mannose. The complex crystallizes in space group P2(1)2(1)2 with a = 70.73, b = 92.97 and C = 47.07A. Synchrotron X-ray data have been measured at 1.86A resolution and the crystal structure of the complex will he solved by the molecular replacement method.
[Back to top] Crystallization and X-Ray Diffraction Data Analysis of Oxyhemoglobin-I from the Catfish Liposarcus anisitsi (PISCES). Smarra, A.L.S., Arni, R.K., De Azevedo, Jr., W.F. Colombo, M.F., and Bonilla-Rodriguez, G. 0.
Hemoglobin remains, despite the enormous amount of research involving this molecule, as a prototype for allosteric models and new conformations. Functional studies carried out on Hemoglobin-I from the South-American Catfish Liposarcus anisitsi suggest the existence of conformational states beyond those already described for human hemoglobin, which could be confirmed crystallographically. The present work represents he initial steps towards that goal.