Protein and Peptide Letters, Vol. 7, No. 6, 2000
Synthesis
of Peptide with Multiple Cysteic Acids. Pp. 359-364.
Anatol
Arendt, J. Hugh McDowell, W. Clay Smith and Paul A. Hargrave
Preparation
of Protected Peptides by Gel-Phase Synthesis on Butanediol Dimethacrylate
Cross-Linked Polystyrene Support. Pp. 365-372.
M. Roice and
V.N. Rajasekharan Pillai
Synthesis
of [ARG 14,17,19,20] Deacetyl-Thymosin a1 and Its Immunological Effect on
the Impaired T-Lymphocytes of Uremic Patients. Pp. 373-379.
T. Abiko and
S. Nakatsubo
Isolation
and Characterization of Novel Neurotoxins from South American Rattlesnake Crotalus
Durissus Terrificus: Optimization of Chromatographic Procedure. Pp.
381-388.
Marcos Hikari
Toyama, Gildo B. Leite, Léa Rodrigues-Simioni, O.S. Saraguacy Hernandez, José
Camillo. Novello and Sergio Marangoni
Physicochemical
Properties and Behavior in Solution of Three Cellulases from Haliotis
fulgens. Pp. 389-396.
Alejandra Hernández-Santoyo,
Arturo Rojo-Domínguez, Enrique García-Hernández and Adela Rodríguez-Romero
Stability
of a Black Eyed Pea Trypsin/Chymotrypsin Inhibitor (BTCI). Pp. 397-401.
Luciano
Paulino Silva, José Roberto S.A. Leite, Carlos Bloch Jr. and Sonia Maria de
Freitas
Molecular Modeling of Kappa Opioid Receptor-Gi Protein Recognition. Pp. 403-409.
Xuhu Wan,
Huang Xiao-Qin, Zhou De-He, Jiang Hua-Liang, Feng Ya-Ping, Chen Kai-Xian, and
Chi Zhi-Qiang
Understanding the Contribution of Individual Tryptophan Residues to Intrinsic Lysozyme Fluorescence. Pp. 411-420.
Irina M.
Kuznetsova, Alexander G. Biktashev, Lubov N. Malova, Natalia A. Bushmarina,
Valdimir N. Uversky and Konstantin K. Turoverov
Crystallization and Preliminary X-Ray Diffraction Studies of BNSP-6, a Myotoxic Phospholipase A2 from Bothrops Neuwiedi Venom. Pp. 421-425.
Marcos R.M.
Fontes, Luiz E. Monteiro, Andreimar M. Soares, Veridiana M. Rodrigues, Reinaldo
J. da Silva and José R. Giglio
[Back to top] Temporin Antibiotic Peptides: A Review and Derivation of a Consensus Sequence.
The temporins are a group of small, linear, basic, highly hydrophobic, antibiotic, peptide amides that were originally isolated from the skin of the European red frog, Rana temporaria. During the past decade, 30 temporin or temporin-like peptides have been isolated from the skins of Anurans. This article presents a brief review of the temporin literature, an analysis of the amino acid sequences of temporins and temporin-like peptides, and the derivation of a composite or consensus sequence, [FLP(I/L)IASLL(S/G)KLL-am].
[Back to top] Synthesis of Peptide with Multiple Cysteic Acids.
Anatol Arendt, J. Hugh McDowell, W. Clay Smith and Paul
A. Hargrave
We have synthesized and purified a 19-amino acid peptide whose sequence is derived from the carboxyl-terminal region of bovine rhodopsin, in which the seven serines and threonines that would normally become phosphorylated have been substituted with cysteic acid. The peptide was synthesized by the Fmoc (DCC/HOBt)+ procedure usingFmoc-(S-4-methoxytrityl) Cys {Fmoc-Cys(Mmt)-OH}. The Mmt group was removed and the resin-bound peptide was oxidized with performic acid, which also removed other blocking groups and cleaved the peptide from the resin.
[Back to top] Preparation of Protected Peptides by Gel-Phase Synthesis on Butanediol Dimethacrylate Cross-Linked Polystyrene Support.
M. Roice & V.N. Rajasekharan Pillai
Preparation of fully protected peptide C-terminal esters in high yield and purity by making use of gel-phase synthesis on chloromethyl butanediol dimethacrylate (BDODMA) crosslinked polystyrene is described. The C-terminal amino acid of the peptide was incorporated by cesium salt method and the step-wise synthesis was carried out using HOBt active ester coupling procedure. The protected peptides were cleaved from the support by trans-esterification. The crude peptides were purified by HPLC and characterized by amino acid analysis, tlc and MALDI TOF MS.
[Back to top] Synthesis of [ARG 14,17,19,20] Deacetyl-Thymosin a1 and Its Immunological Effect on the Impaired T-Lymphocytes of Uremic Patients.
T. Abiko and S. Nakatsubo
A deacetyl-thymosin a1 analogue containing four arginines at positions 14, 17, 19, and 20, instead of four lysines was synthesized by the manual solid-phase method. The synthetic [Arg 14,17,19,20] deacetyl-thymosin a1 and deacetyl-thymosin a1 were tested for the effect on impaired T-lymphocyte transformation by phytohemagglutinin in uremic patients suffering from recurrent infectious diseases. The restoring activity on the impaired phytohemagglutinin stimulation of T-lymphocytes was obtained after incubation of peripheral lymphocytes isolated from uremic patients with the synthetic [Arg 14,17,19,20] deacetyl-thymosin a1. This analogue exhibited a far stronger restoring effect than that of our synthetic deacetyl-thymosin a1.
[Back to top] Isolation and Characterization of Novel Neurotoxins from South American Rattlesnake Crotalus Durissus Terrificus: Optimization of Chromatographic Procedure.
Marcos Hikari Toyama, Gildo B. Leite, Léa
Rodrigues-Simioni, O.S. Saraguacy Hernandez, José Camillo. Novello and Sergio
Marangoni
Molecular exclusion HPLC chromatography on a Protein Pak SW 300 column and reversed phase HPLC on a m-Bondapack C18 column were used to purify four novel neurotoxins from the crotoxin complex, and five crotoxin subunits (PLA2 and crotapotin) as well as two crotamine isoforms. A single preparative reversed phase HPLC steps yielded a major basic myotoxin followed by the subunits of crotoxin (three PLA2 isoforms and two crotapotin isoforms). This modified procedure may be useful for the rapid, large-scale purification of toxins from Crotalid venoms.
[Back to top] Physicochemical Properties and Behavior in Solution of Three Cellulases from Haliotis fulgens.
Alejandra Hernández-Santoyo, Arturo Rojo-Domínguez,
Enrique García-Hernández and Adela Rodríguez-Romero
Three proteins that showed activity over cellulosic substrates were isolated from the hepatopancreas of the blue abalone Haliotis fulgens. The purified cellulases are acidic with molecular weights between 17 900 and 30 300. Activity experiments indicated that they are endo-, exo- and b-glucanases. These proteins form large aggregates as demonstrated by means of DLS experiments, which can be dissociated in the presence of polyethylene glycol and mannitol. The circular dichroism spectra in the far UV indicated that the enzymes belong to the b-b family.
[Back to top] Stability of a Black Eyed Pea Trypsin/Chymotrypsin Inhibitor (BTCI).
Luciano Paulino Silva, José Roberto S.A. Leite, Carlos
Bloch Jr. and Sonia Maria de Freitas
The stability of BTCI has been investigated as function of pH and temperature, following its inhibitory activity against trypsin. The isolated inhibitor of 9, 084 Da is stable over pH 3 to 12 at 25oC. BTCI showed high thermal stability ranging from 25 to 95oC. At pH 3 and 7. However, the protein lost about 20% of its inhibitory activity over 75oC at pH 8.2. The results indicated that BTCI is extremely stable to heat and pH as typical of Bowman-Birk inhibitors.
[Back to top] Molecular Modeling of Kappa Opioid Receptor-Gi Protein Recognition.
Xuhu Wan, Huang Xiao-Qin, Zhou De-He, Jiang Hua-Liang,
Feng Ya-Ping, Chen Kai-Xian, and Chi Zhi-Qiang
This work theoretically studied the mechanism involved in recognition of Gi protein and kappa opioid receptor with shae matching method. We used human kappa opioid receptor model docking with dynorphin A(1-8)(HKOR_DYN) to mimic the active form of HKOR. The modeling results suggested the cluster formed by A4, A5, AG and B6 in the alpha subunit of Gi protein (Gai) might be the binding core with intercellular solvent face of HKOR and some residues of the intracellular loops play an important role in Gi protein coupling.
[Back to top] Understanding the Contribution of Individual Tryptophan Residues to Intrinsic Lysozyme Fluorescence.
Irina M. Kuznetsova, Alexander G. Biktashev, Lubov N.
Malova, Natalia A. Bushmarina, Valdimir N. Uversky and Konstantin K. Turoverov
Packing density and other properties of the microenvironment of tryptophan residues in hen egg-white lysozyme macromolecule have been studied on the basis of the known 3D structure of this protein. Results presented here suggest that the efficiency of action of the group which, in principal, can effect tryptophan fluorescence depends not only on the distance between this group and the indole ring, but to a great extent upon the location of this group relative to the indole ring.
[Back to top] Crystallization and Preliminary X-Ray Diffraction Studies of BNSP-6, A Myotoxic Phospholipase A2 from Bothrops Neuwiedi Venom.
Marcos R.M. Fontes, Luiz E. Monteiro, Andreimar M.
Soares, Veridiana M. Rodrigues, Reinaldo J. da Silva and José R. Giglio
BnSP-6 (myotoxin I) is a phospholipase A2 homologue isolated from Bothrops neuwiedi pauloensis venom. Crystals of BnSP-6 were obtained which diffracted X-rays to 2.5 Å resolution using a synchrotron radiation source at room temperature and belong to space group P3121. The unit cell dimensions are a=b=57.7, c=131.1 Å. The structure was solved by molecular replacement using the coordinates of botropstoxin I from B. jararacussu venom. There are two molecules in the asymmetric unit.