Amino Acid
Sequence and Molecular Modelling of A Lipid Transfer Protein from Sunflower (Helianthus
Annus L.) Seeds Pp.241-248
Effect of
Lyotropic Salts on the Stability of a Subtilisin-Like Proteinase From a
Psychrotrophic Vibrio-Species, Proteinase K and Aqualysin I Pp.249-255
Epitope-Paratope Recognition by Knowledge Based Correlation Mapping Using Hopfield Network Pp.257-264
One-Step
Affinity Purification of Human Mu-Opioid Receptor Overexpressed in Baculovirus
System Pp.265-272
The Native Gene of Anti-Lps Factor From Tachypleus Tridentatus: Cloning, Expression and Its Bacteriostatic Activity in Vitro Pp.273-280
Design
of Dermaseptin S3 Analogues Having Bacterial Cell Selectivity Without Mammalian
Cell Toxicity Pp.281-288
Chemical
Syntheses of cart (Human, 55-102) Nalogues and their Anorectic Effect on Food
Intake in Rats Induced by Neuropeptidey Y Pp.289-295
Secondary
Structure of the Homologous Proteins, A a-Fetoprotein
And Serum Albumin, From Their Circular Dichroism and Infrared Spectra Pp.297-302
Galactosylation
Thermodynamics of E. Coli B b-Galactosidase
by ONPG and PNPG Pp.303-306
Conformation
of Di-N-Propyl Glycine Residue in A Glycine Rich Peptide Sequence Pp.307-315
Crystallization
and Preliminary X-Ray Crystallographic Studies of Human Autocrine Motility
Factor Pp.317-322
Crystallization
and X-Ray Data Analysis of the Extended Dna-Binding Domain of the E2 Bovine
Papillomavirus Type 1 Protein Pp.323-326
Charcterization
and Preliminary X-Ray Diffraction Analysis of the Mutant Pro229ser of
Thermostable Catechol 2,3-Dioxygenase Pp.327-332
Abstracts
[Back to top] Amino Acid Sequence
and Molecular Modelling of A Lipid Transfer Protein from Sunflower (Helianthus
Annus L.) Seeds
Suzanne Luckett , Richard B. Sessions
, Louise Michaelson , Michael J.
Naldrett , Anthony R. Clarke ,
Roger Fido , Arthur S. Tatham and Peter R. Shewry
The major lipid transfer
protein has been purified from sunflower seeds and its amino acid sequence
determined. A homology model has been built based on the rice LTP, which
suggests that the protein can bind palmitic acid. The sequence may therefore
correspond to previously characterized proteins with lipid transfer and
antifungal activities.
[Back to top] Effect Of Lyotropic Salts on the Stability
of A Subtilisin-Like Proteinase From A Psychrotrophic Vibrio-Species,
Proteinase K and Aqualysin I
Magnús M. Kristjánsson and Ólafur
Th. Magnússon
The stability of three related subtilisin -like proteinases; a proteinase from a psychrotrophic Vibrio-species, proteinase K and aqualysin I from the thermophile Thermus aquaticus, to denaturation by GdmSCN, was measured in the presence and absence of selected lyotropic salts (sulfate and thiocyanate). How these salts affect protein stability has been ascribed to their effect on the strength of hydrophobic interactions. The salts affected the stability of the proteinases according to their ranking in the lyotropic salt series. The effect of the salts was smallest, however, on the stability of the cold-adapted Vibrio-proteinase, but largest for the thermophilic aqualysin I. These results suggest that the cold-adapted enzyme may be less dependent on hydrophobic interactions for structural stability than its counterparts adapted to higher temperatures.
[Back to top] Epitope-Paratope Recognition by
Knowledge Based Correlation Mapping Using Hopfield Network
Rajani R. Joshi
The knowledge based correlation mapping (KBCM) is devised here as a unified measure of the information on antigen-antibody complexes available in the Protein Data Banks. This mapping is estimated from a random training sample using a Hopfield Network while representing the epitope and paratope in a complex as 3-dimensional attributed graphs. Identification of paratopes on immunoglobulins against given epitopes is carried out using pattern ‘matching’ on Hopfield Network with the help of the KBCM.
[Back to top] One-Step Affinity Purification of Human Mu-Opioid Receptor
Overexpressed in Baculovirus System
Chen Li-Wei, Feng Ya-Ping, Zhou De-He
, Xu Xue -Jun, Chen Jie, Wei Qiang, and Chi Zhi-Qiang
Affinity chromatography on Ni-nitrilotriacetic acid agarose was used to purify human mu-opioid receptors overexpressed in Sf9 cells. We introduced a tag of 6 consecutive histidines at its carboxyl terminus for easy purification of recombinant proteins. The binding activity and identification of the purified receptor were determined by receptor binding assay, SDS-PAGE and Western blotting analysis. This procedure used in the paper offer an easy and fast route to the purification of recombinant human mu-opioid receptors.
[Back to top] The Native Gene of Anti-Lps Factor From Tachypleus
Tridentatus: Cloning, Expression and Its Bacteriostatic Activity in Vitro
Dong-Ning Wang , Lin Chen , Jie-Wu Liu , Zhi-Yong He , Wei-Jie
Zhang , Xiang-Fu Wu
The native gene information of anti-LPS factor from Tachypleus tridentatus was first reported in this study. The total RNA was extracted from amebocytes of Tachypleus tridentatus and the first-strand cDNA was synthesized by RT-PCR. Then the amplified cDNA of TALF was cloned and expressed in Pichia pastoris. The biological activity of the recombinant proteins obtained have been confirmed by LPS neutralization assay and bacteriostatic assay in vitro.
[Back to top] Design of Dermaseptin S3 Analogues Having Bacterial Cell Selectivity Without Mammalian Cell Toxicity
Song
Yub Shin, Sung-Tae Yang, Soo Hyun Eom, Woo Keun Song, Yangmee Kim,Kyung-Soo
Hahm & Jae Il Kim,
Dermaseptin S3 (DS3) is a a-helical antimicrobial peptide isolated from the South American frogs Phyllomedusa sauvagii. Insertion of Gly-Pro sequence into DS3 resulted in a drastic reduction in the mammalian cell toxicity against human red blood and normal fibroblast cells, with retention of the antibacterial activity. The results indicated that the bend created by insertion of Gly-Pro in the continuous a-helical peptide plays an important role in providing high selectivity against bacterial cells without mammalian cell toxicity.
[Back to top] Chemical Syntheses of cart (Human, 55-102)
Nalogues and their Anorectic Effect on Food Intake in Rats Induced by
Neuropeptidey
To understand the comparative effect on the anorectic property of the replacement of two Tyr residues at positions 58 and 62 in CART(55-102) by two Tyr(Me) or Phe(NO2) residues, [Tyr(Me)58,62 ]CART55-102 and [Phe(NO2)58,62 ]CART55-102 were synthesized. In vivo inhibitory activity on the food intake induced by NPY was compared with that of the parent peptide. [Phe(NO2)58,62 ]CART55-62 exhibited 2-fold higher anorectic activity on the food intake induced by NPY as compared with the parent peptide. [Tyr(Me)58,62 ]CART55-102 exhibited less potent anorectic activity than the parent peptide. These results indicate that aromaticity of two Tyr residues in CART55-102 is important for anorectic activity on the food intake induced by NPY and an electrophilic group on aromatic rings such as -NO2 enhanced anorectic activity more than that of the parent peptide. On the contrary, an electron-donating group on aromatic rings (Tyr 58,62 ) such as CH3O- influenced to reduce anorectic activity as compared with the parent peptide.
[Back to top] Secondary Structure of the Homologous
Proteins, A A-Fetoprotein And Serum Albumin, From Their Circular Dichroism and
Infrared Spectra
Secondary structure composition of two homologous proteins, a-fetoprotein and serum albumin, has been compared using circular dichroism and Fourier-transform infrared spectroscopy. CD-only analysis gives an accurate a-helical content for SA, but underestimates the predicted value for AFP. In contrast, FTIR-only analysis underestimates the helical content of SA. However, analysis of hybrid IR-CD spectra gives more accurate values for secondary structure composition for both AFP and SA and shows that the secondary structures of these proteins are the same.
[Back to top] Galactosylation Thermodynamics of E. Coli
B b-Galactosidase by ONPG and PNPG
Michaelis-Menten analysis of the hydrolyses of ONPG and PNPG by E. coli b-galactosidase were performed from 5.5 to 45°C. Analysis of the T-dependence of KM and kcat reveals the thermodynamics for formation of the E/S complex and attainment of the galactosylation transition state, respectively. While the binding and transition state free energies are similar for each substrate, the enthalpic and entropic contributions are found to differ substantially.
[Back to top] Conformation of Di-N-Propyl Glycine
Residue in A Glycine Rich Peptide Sequence
The peptide Boc-(Gly-Dpg-Gly)3-OBzl (I) has been synthesized to examine the conformational preferences of Dpg residues in a peptide sequence rich in glycine, which is known to be a poor helix promoter. The detailed NMR studies revealed that peptide (I) adopts a helical conformation in CDCl3.
[Back to top] Crystallization and Preliminary X-Ray
Crystallographic Studies of Human Autocrine Motility Factor Hiroshi
Uemura , Nobutada Tanaka , Tatsuyoshi
Funasaka , Arayo Haga , Hisamitsu
Nagase , Avraham Raz , and Kazuo T. Nakamura
Human autocrine motility factor (hAMF), a tumor-secreted cytokine which stimulates cell migration in vitro and metastasis in vivo, has been crystallized by the hanging-drop vapour diffusion method. The crystals belong to an orthorhombic space group P212121 with the cell dimensions of a = 80.79 Å, b = 107.1 Å, and c = 270.9 Å. There are two dimers per asymmetric unit. The crystals diffract to at least 2.0 Å resolution and are suitable for X-ray structure analysis at high resolution.
[Back to top] Crystallization and
X-Ray Data Analysis of the Extended Dna-Binding Domain of the E2 Bovine
Papillomavirus Type 1 Protein
Hanging-drop vapour-diffusion method was used for the crystallization of the extended DNA-binding domain (residues 309-410) of the E2 protein from Bovine Papillomavirus Type 1. X-ray data collection at 2.0 Å resolutions was performed using synchrotron radiation. The crystal symmetry could be described by the space group P3121 and with unit cell parameters a = b = 55.3 Å, c = 203.4 Å. The protein structure was solved by molecular replacement.
[Back to top] Charcterization and
Preliminary X-Ray Diffraction Analysis of the Mutant Pro229ser of Thermostable
Catechol 2,3-Dioxygenase
The thermostability
of the mutant Pro229Ser of thermostable catechol 2,3-dioxygenase was decreased
about 10.2 °C compared with that of the wild type enzyme. By the hanging-drop
vapor-diffusion method using PEG 400 as a precipitant, Pro229Ser was
crystallized at 4 °C. The crystals belonged to the orthorhombic space group I222 with
unit-cell parameters a = 70.59 Å, b = 74.15 Å, c = 133.11 Å (a =b=g=900). The Matthews parameter (Vm) was
calculated to be 2.49 Å 3 D -1, and the solvent content was about 51 %.