Protein & Peptide Letters, Volume 10, No. 2, 2003

Protein & Peptide Letters, Volume 10, No. 2, 2003

Contents

Mini - Review

[Abstract] Normal Mode Analysis with Simplified Models to Investigate the Global Dynamics of Biological Systems Pp.119-132

Florence Tama

Regular Papers

[Abstract] A Simplified Model of the Binding Interaction Between Stromal Cell-Derived Factor-1 Chemokine and Cxc Chemokine Receptor 4 Pp.133-138

Pasquale Palladino , Carlo Pedone , Raffaele Ragone , Filomena Rossi , Michele Saviano , and Ettore Benedetti

[Abstract] Binding Energetics of the Inhibitor Cystatin to the Cysteine Proteinase Actinidin Pp.139-145

Maricela Neria-Rios, Jaqueline Padilla-Zuniga, Enrique Garcia-Hernandez, Salvador R. Tello-Solis and Rafael A. Zubillaga

[Abstract] Synthesis of Chemotactic Peptide Analogs with a Photoaffinity Cross-Linker As New Probes for Analysis of Receptor-Ligand Interaction Pp.147-153

Mitsukuni Shibue , Masahiro Yoshiki , Masaya Miyazaki , Ichiro Fujita , Satoshi Osada , Seiji Yasuda, Hiroyuki Nakamura, Yuhei Hamasaki , Michio Kondo , Hideaki Maeda , Hiroaki Kodama and Hazime Shimizu

[Abstract] Investigations on the Binding of Mercury Ions to Albumins Employing Differential Pulse Voltammetry Pp.155-164

Clarissa Silva Pires de Castro1, Jurandir Rodrigues SouzaDe2 and Carlos Bloch Jr.

[Abstract] Purification and Characterization of a Lectin from Annona Coriacea Seeds Pp.165-173

Mirela B. Coelho , Maria das Graças M. Freire , Marcos H. Toyama , Sergio Marangoni , Jose C. Novello and Maria Ligia R. Macedo

[Abstract] An Efficient Fusion Expression System for Protein and Peptide Overexpression in Escherichia Coli and Nmr Sample Preparation Pp.175-181

Yuan Cheng, Dongsheng Liu, Yanming Feng, Guozhong Jing

[Abstract] Isolation and Some Characterizations of a Glycosylated Fibrinolytic Enzyme of Earthworm, Eisenia Fetida Pp.183-190

Li Li, Jing Zhao and Rong-Qiao He

[Abstract] Renal Alterations Promoted by the Lectins from Canavalia Ensiformis (Cona) and Dioclea Guianensis (Dguil) Seeds Pp.191-197

Alexandre Havt , Paulo S. F. Barbosa, Ticiana M. Sousa, Alice M. C. Martins, Arlandia C. L. Nobre, Kyria S. Nascimento , Edson H. Teixeira, Vicente P. T. Pinto, Alexandre H. Sampaio, Manassés C. Fonteles , Benildo S. Cavada , Helena S. A. Monteiro

[Abstract] The Effect of Polyols on the Reactivation of Guanidium Chloride-Denatured Arginine Kinase from Shrimp Feneropenaeus Chinensis Muscle Pp.199-211

Zhenhang Yu and Biao Li

[Abstract] Expression of Recombinant Xvax2 Peptide-104 of Xenopus Laevis and Preparation of the Antibody Pp.213-219

Yang Liu , Ying Liu , Wei Liu , Giuseppe Lupo , Rong-qiao He and Giuseppina Barsacchi

Crystallization Report

[Abstract] Crystallization of the N-Terminal Domain of Dff45: The Mutual Chaperone Mechanism is

Challenged Pp.221-225

Li T. , Li X. , Yang W. , Rao Z. & Zhai Zh.

Abstracts

[Back to top] Normal Mode Analysis with Simplified Models to Investigate the Global Dynamics of Biological Systems

Florence Tama

Dynamical properties of macromolecules are increasingly being recognized as significantly contributing to biological functions, including catalysis, regulation of activity, etc. In this review, theoretical approaches to the study of dynamics of biological systems and their application are discussed. In particular, simplified models for the normal mode analysis are described.

[Back to top] A Simplified Model of the Binding Interaction Between Stromal Cell-Derived Factor-1 Chemokine and Cxc Chemokine Receptor 4

Pasquale Palladino , Carlo Pedone , Raffaele Ragone , Filomena Rossi , Michele Saviano , and Ettore Benedetti

We have synthesised the CXC chemokine receptor 4 (29-39) peptide, CXCR4{29-39}. This peptide is located in the N-terminal region of the receptor and is likely to be involved in the docking step of the receptor interaction with its natural ligand stromal cell-derived factor-1 chemokine, SDF-1. Preliminary experiments, performed in the presence of micellar detergents to model a membrane-like environment, show that the (1-17) segment of SDF-1 binds to CXCR4{29-39}.

[Back to top] Binding Energetics of the Inhibitor Cystatin to the Cysteine Proteinase Actinidin

Maricela Neria-Rios, Jaqueline Padilla-Zuniga, Enrique Garcia-Hernandez, Salvador R. Tello-Solis and Rafael A. Zubillaga

The binding energetics of actinidin to chicken cystatin was determined from fluorometric titrations at different temperatures. It is shown that the association of actinidin with cystatin is both enthalpically and entropically driven, with a negative change in the heat capacity. The molecular basis of these contributions are analyzed within the framework of surface-area models, using a 3D model of the actinidin-cystatin complex, which was obtained using the x-ray structure of the homologous complex papain-stefin B as template.

[Back to top] Synthesis of Chemotactic Peptide Analogs with a Photoaffinity Cross-Linker as New Probes for Analysis of Receptor-Ligand Interaction

Formylpeptide receptors are well-characterized receptors which participate in host defense responses of neutrophils. We designed and synthesized chemotactic peptide analog with pbenzoylphenylalanine (Bpa) and biotin to probe structural and mechanistic aspects of peptide-receptor interaction. These peptides possess biological activities which were dependent upon spacer residue length of and Bpa position. The covalent photoaffinity label was detected by Streptavidine-blot, which was inhibited by the parent peptide.

[Back to top] Investigations on The Binding of Mercury Ions to Albumins Employing Differential Pulse Voltammetry

Clarissa Silva Pires de Castro, Jurandir Rodrigues SouzaDe and Carlos Bloch Jr

Binding of mercury to BSA and Ovalbumin was investigated by Differential Pulse Voltammetry. The method relies on the direct monitoring of peak current variation due to mercury oxidation in the presence of these two albumins. Linear calibration graphs were obtained for both BSA and Ovalbumin in concentrations ranging from 2.49 x 10^-7 to 19.6 x 10^-9 mol L^-1. The acquired data was used to quantify these two proteins independently and to calculate the dissociation constants of Hg-BSA and Hg-Ovalbumin complexes.

[Back to top] Purification and Characterization of A Lectin from Annona Coriacea Seeds

Mirela B. Coelho , Maria das Graças M. Freire , Marcos H. Toyama , Sergio Marangoni , Jose C. Novello and Maria Ligia R. Macedo

A novel lectin, denominated ACLEC, was isolated from Annona coriacea seeds, belonging to the Annonaceae family. The lectin presented one protein band in SDS-PAGE of 14 kDa. Of the sugars tested, Dglucose and D-mannose were the best inhibitors. A search sequence database showed that ACLEC had homology with other plant lectins, belonging to leguminous lectin family.

[Back to top] An Efficient Fusion Expression System for Protein and Peptide Overexpression in Escherichia Coli and Nmr Sample Preparation

Yuan Cheng, Dongsheng Liu, Yanming Feng, Guozhong Jing

An efficient fusion expression system with a small fusion partner, His6-tagged N-terminal fragment of staphylococcal nuclease R, has been constructed and tested with two genes. The results show that the system is not only suitable for overexpression of small proteins and peptides but simplifies purification of target proteins and peptides. The study also provides a practical method for preparation of isotope-labeled protein sample for NMR analysis.

[Back to top] Isolation and Some Characterizations of A Glycosylated Fibrinolytic Enzyme of Earthworm, Eisenia Fetida

Li Li, Jing Zhao and Rong-Qiao He

Resin coupled with m-aminophenylbornic acid was used to isolate a glycosylated component from homogenate of earthworm (Eisenia fetida). The fraction showed a single band on SDS-PAGE with a molecular weight of 34,193 Da determined by mass spectroscopy. The N-terminal region is AQVCCPDI, different from those of earthworm fibrinolytic enzymes reported previously (Nakajima et al. 1993). This glycosylated component showed an activity on digesting both Chromozym-TH and fibrin, suggesting that it is a novel fibrinolytic enzyme.

[Back to top] Renal Alterations Promoted by the Lectins from Canavalia Ensiformis (Cona) and Dioclea Guianensis (Dguil) Seeds

The lectin from the seeds of Canavalia ensiformis (ConA) and Dioclea guianensis (DguiL) was tested upon its renal effects using the isolated perfusion rat kidney method. Both lectins (10 mg/ml) affected perfusion pressure and renal vascular resistance, but DguiL showed a much greater action than ConA. However, ConA, but not DguiL, affected potassium tubular transport.

[Back to top] The Effect of Polyols on the Reactivation of Guanidium Chloride-Denatured Arginine Kinase from Shrimp Feneropenaeus Chinensis Muscle

Zhenhang Yu and Biao Li

The influence of glycerol, sorbitol, glucose and sucrose, on the refolding and reactivation courses of guanidine-denatured arginine kinase was studied. Glycerol, sucrose, and sorbitol, but not glucose, could improve the reactivation of the denatured arginine kinase, although in all cases aggregation was inhibited. Size exclusion chromatography showed that misfolded products were still formed during polyol-assisted refolding. The chemical and physical characteristics of polyols might explain the various observations.

[Back to top] Expression of Recombinant Xvax2 Peptide-104 of Xenopus Laevis and Preparation of the Antibody

Yang Liu , Ying Liu , Wei Liu , Giuseppe Lupo , Rong-qiao He and Giuseppina Barsacchi

An N-terminal polypeptide (XVAX2-P104) of the XVAX2 protein, which is involved in controlling the dorsoventral patterning of the retina in Xenopus laevis, was expressed and purified as a Histagged fusion protein (pHis-XVAX2-P104), and it was employed to generate an anti-XVAX2 antibody in New Zealand white rabbits. ELISA analysis shows that the titer of the antibody is as high as ~1:100,000. The antibody could specifically recognize the full-length XVAX2 protein in extracts of Xenopus embryos, as determined by Western Blotting.

[Back to top] Crystallization of the N-Terminal Domain of Dff45: The Mutual Chaperone Mechanism is Challenged

Li T. , Li X. , Yang W. , Rao Z. & Zhai Zh.

DNA fragmentation factor 45 (DFF45) regulates DNase DFF40 as its inhibitor and chaperone. It was reported that the N-terminal domain (NTD) of DFF45 alone is disordered and DFF40 is necessary as a mutual chaperone for the folding of NTD. However, here we reported the crystallization of DFF45 NTD. These crystals diffract to 9Å using a synchrotron radiation source. In spite of the low resolution, the demonstration of crystal formation indicates that DFF45 NTD itself is not unstructured, which strongly questions the mutual chaperone speculation about DFF45 and DFF40.