Diversity of Equilibrium Compact Forms of Denatured Globular Proteins. Pp. 355-368.
Vladimir N. Uversky
[Abstract]
Papers
Simulation of The Unfolding of a Leucine Zipper. Pp. 369-374.
Karen Flanagan and Julia M. Goodfellow
[Abstract]
Identification of an Esterase from Bacillus acidocaldarius with Sequence
Similarity to a Hormone Sensitive Lipase Subfamily. Pp. 375-382.
Giuseppe Manco, Elena Adinolfi, Francesca M. Pisani, Vitale Carratore and
Mosé Rossi
[Abstract]
Bactericidal Properties of HIV- 1 VPR C-Terminal Sequences. Pp. 383-390.
P. Failla, L.A. Castelli, A.A. Azad and L G. Macreadie
[Abstract]
High-Level Expression and Efficient Purification of the Antimicrobial Peptide
Gaegurin 4 in E. coli. Pp. 391-396.
Jiyoung Kim, Jin Mo Park and Byeong Jae Lee
[Abstract]
The Influence of N-Terminal Fusion Partners on the Production and Folding of
Recombinant Human Procathepsin E. Pp. 397-404.
Peter J. Tatnell and John Kay
[Abstract]
Synthetic Peptide as Inhibitors of Human Complement Activation. Pp. 405-408.
Aruna Kapil, Bijoy Kundu, Sanjay K. Khare and Manisha Shukla
[Abstract]
Assay of Lumbrokinase with a Chromophoric Substrate. Pp. 409-414.
Jing Zhou, Rong Fan, Chen Wu and Rong-Qiao He
[Abstract]
Crystallisation Report
Purification and Crystallization of A 2,3-Dihydroxybiphenyl Dioxygenase ETBC from Rhodococcus Sp. Strain RHA1. Pp. 415-418.
Keisuke Sugimoto, Takahiro Yamada, James F. Hauschild, Toshiya Senda, Eiji Masai, Masao Fukuda and Yukio Mitsui
[Abstract]
[Back to top] Diversity of Equilibrium Compact Forms of Denatured Globular Proteins. Vladimir N. Uversky.
The problem of the variety of the equilibrium denatured forms of globular proteins is considered. The crucial observation is emphasized that the molten globule is not the only equilibrium intermediate of a given protein. The structural properties of a protein molecule in different intermediate conformations are described. An important fact is emphasized that among several compact denatured forms only the molten globule represents a distinct phase state of a protein molecule.
[Back to top] Simulation of The Unfolding of a Leucine Zipper. Karen Flanagan and Julia M. Goodfellow.
Molecular dynamics simulations of the 33 residue coiled-coil domain of the GCN4 leucine zipper GCN4-p1 have been carried out at 298 and 328K. The room temperature simulation correlated well with the findings of Rozelle et al. [1] and is consistent with NMR data [2]. The higher temperature simulation indicates the presence of a conformational substate on the folding pathway. Analysis of breaking of hydrogen bonds and hydrophobic interactions gives information on the order of protein unfolding events.
[Back to top] Identification of an Esterase from Bacillus acidocaldarius with Sequence Similarity to a Hormone Sensitive Lipase Subfamily. Giuseppe Manco, Elena Adinolfi, Francesca M. Pisani, Vitale Carratore and Mosé Rossi.
We purified an esterase from Bacillus acidocaldarius whose NH2-terminal sequence corresponds to an open reading frame (ORF) previously reported to show homology with the mammalian hormone sensitive lipase (HSL)-like subgroup of the esterase/lipase family. The enzyme displays esterase but not lipase activity. The enzyme we predicted the alpha/beta-hydrolase topological fold common to several esterases and lipases and identified Ser 155, Asp 252 and His 282 as the putative members of the catalytic triad.
[Back to top] Bactericidal Properties of HIV-1 VPR C-Terminal Sequences. P. Failla, L.A. Castelli, A.A. Azad and L G. Macreadie.
Vpr is a virion-associated protein of human immunodeficiency virus type 1 (HIV-1). Vpr is toxic to human and yeast cells when produced in the cell or when added externally, due to the presence of the sequence HFRIGCRHSRIG, which represent Vpr residues 71-82. Here we show that the C-terminal sequence of Vpr also causes toxicity in E. coli. The intracellular but not the extracellular toxicity can be partially overcome with the selection of a suppressor of Vpr toxicity (svt) mutant E. coli strain.
[Back to top] High-Level Expression and Efficient Purification of the Antimicrobial Peptide Gaegurin 4 in E. coli. Jiyoung Kim, Jin Mo Park and Byeong Jae Lee.
Gaegurin 4 (GGN4), an antimicrobial peptide isolated from Rana rugosa skin, was expressed in Escherichia coli as a form of inactive glutathione S-transferase (GST) fusion polypeptide. Recombinant GGN4 was obtained by enzymatic and chemical processing of the fusion protein and purification of the processed products by affinity chromatography and high performance liquid chromatography (HPLC). 2.2 mg of biologically active recombinant GGN4 was obtained from 7 liters of E. coli culture. This strategy may be applied to preparation of other recombinant peptides which cause host cell toxicity.
[Back to top] The Influence of N-Terminal Fusion Partners on the Production and Folding of Recombinant Human Procathepsin E. Peter J. Tatnell and John Kay.
The production of human procathepsin E in Escherichia coli was optimised by expression in the form of N-terminal tagged fusion proteins using several T7-RNA polymerase dependent pET-vectors. The nature of the N-terminal fusion partner did not significantly influence the amount of the precursor that accumulated. However, the yield of active, mature cathepsin E that was recovered after refolding varied by as much as ten-fold.
[Back to top] Synthetic Peptide as Inhibitors of Human Complement Activation. Aruna Kapil, Bijoy Kundu, Sanjay K. Khare and Manisha Shukla.
Several structurally diverse hexapeptides related to IgE have been evaluated for their ability to inhibit complement activation by using hemolytic assay. Out of the compounds tested, four hexapeptides exhibited total suppression of human complement.
[Back to top] Assay of Lumbrokinase with a Chromophoric Substrate. Jing Zhou, Rong Fan, Chen Wu and Rong-Qiao He.
A new method to assay lumbrokinase with a chromophoric substrate was introduced. Compared with the fibrin plate method, the chromophoric assay is rapid, convenient, accurate and repeatable. The activity of lumbrokinase can be defined as the international unit directly with this method.
[Back to top] Purification and Crystallization of A 2,3-Dihydroxybiphenyl Dioxygenase ETBC from Rhodococcus Sp. Strain RHA1. Keisuke Sugimoto, Takahiro Yamada, James F. Hauschild, Toshiya Senda, Eiji Masai, Masao Fukuda and Yukio Mitsui.
Crystals have been obtained for a 2,3-dihydroxybiphenyl dioxygenase (conventionally called EtbC) from a polychlorinated biphenyl (PCB)-degrader, Rhodococcus sp. strain RHA1. The crystals were grown using PEG400 as a precipitating agent. The crystals belonged to a tetragonal space group (P4) and diffracted to 2.6A.