Protein & Peptide Letters, Volume 9, No. 6, 2002
Contents
Characterization of Rhodamine Conjugated
Agiotensin II Peptide: Synthesis, Analysis and Receptor Binding and
Internalization. Pp.465-476
Satya
P. Yadav, Sadashiv Karnick, Wei-Zhen Shen, and Jingli Zhang
Virus-Cell Fusion Inhibitory Activity of
Novel Analogue Peptides Based on the HP (2-20) Derived from N-Terminus of
Helicobacter Pylori Ribosomal Protein L1 Pp.477-486
Eun-Rhan
Woo, Dong Gun Lee, Young-Su Chang, Yoonkyung Park and Kyung-Soo Hahm
Structure-Antibacterial Activity Relationship
of Cecropin A Derivatives Pp.487-493
Song
Yub Shin, Keun Woo Lee, Yangmee Kim, Jae II Kim, Kyung-Soo Hahm and Shin-Won
Kang
A Model for the Cytoplasmic Domain of the
Influenza A Virus M2 Channel by Analogy to the HIV-1 Vpu Protein Pp.495-502
José
W. Saldanha, Peter E. Czabotar, Alan J. Hay and William R. Taylor
Selective Precipitation of Haptoglobin and a2-Macroglobulin from Human Serum Using Alocasia macrorhiza Tuber Protein Pp.503-510
B.
Shivananda Nayak, N. Jagadish Ulloor and B. Shivaraj
Biochemical Characterization of Apoptotic Cleavage
of KH-Type Splicing Regulatory Protein (KSRP)/Far Upstream Element-Binding
Protein 2 (FBP2) Pp.511-519
Heeyoung
Seok, Jinsun Cho, Minseok Cheon and II-Seon Park
Acyl-Gelatins for Cell-Hybrid Biomaterials: Preparation
of Gelatins with High Melting Point and Affinity for Hydrophobic Surfaces Pp.521-528
Keiichi
Miyamoto, Hiroko Chinzei and Takashi Komai
Synthesis of Alkyl and Aryl Esters of
N-Protected b-Homoamino Acids
from N-Protected a-Aminodiazoketones Pp.529-532
Antibiotic Properties of Novel Synthetic
Temporin A Analogs and a Cecropin A-Temporin A Hybrid Peptide Pp.533-543
David
Wade, Jan-Ingmar Flock, Charlotta Edlund, Ingegerd Löfving-Arvholm,Matti
Sällberg, Tomas Bergman, Angela Silveira, Cecille Unson,Louise Rollins-Smith,
Jerzy Silberring, Malcom Richardson, Pentti Kuusela and Hilkka Lankinen
A Direct Continuous pH-Spectrophotometric
Assay for Arginine Kinase Activity Pp.545-552
Yu
Z., Pan J. and Zhou H.M.
Crystallization
Report
Purification, Crystallization and Preliminary
X-Ray Studies of Human Augmenter of Liver Regeneration Pp.553-556
Chao-Neng
Ji, Zai-Long Cai, Gen-Tao Cao, Gang Yin, Bing-Hua Jiao Tao Jiang, Guang Shu,
Ji-Fang Mao, Yi Xie, and Yu-Min Mao
[Back to top] Characterization of Rhodamine Conjugated
Agiotensin II Peptide: Synthesis, Analysis and Receptor Binding and
Internalization.
Satya P. Yadav, Sadashiv Karnick, Wei-Zhen Shen, and
Jingli Zhang
The results in this
study show that the rhodamine fluorophore can be specifically conjugated to
Angiotensin II at Lys3 residue (substituted for a Val) without
altering the biological activity of the parent compound. The conjugated peptide
was characterized using HPLC, mass spectrometry, and N-terminal sequencing. The
rhodamine-Angiotensin II binds effectively to AT1 receptor and gets
internalized in clathrin coated vesicles by endocytosis. These results clearly
suggest the usefulness of fluorophore-conjugated peptides in studies such as,
ligand-receptor binding, and ligand-receptor complex internalization, for drug
delivery using cell receptors and as an alternative to peptide hormone
radioimmunoassays.
[Back to top] Virus-Cell Fusion Inhibitory Activity of
Novel Analogue Peptides Based on the HP (2-20) Derived from N-Terminus of
Helicobacter Pylori Ribosomal Protein L1
Eun-Rhan Woo, Dong Gun Lee, Young-Su Chang, Yoonkyung Park and Kyung-Soo Hahm
HP (2-20) (AKKVFKRLEKLFSKIQNDK) is the antibacterial sequence derived from N-terminus of Helicobacter pylori Ribosomal Protein L1 (RPL1). It has a broad-spectrum microbicidal activity in vitro that is thought to be related to the membrane-disruptive properties of the peptide. Based on the putative membrane-targeted mode of action, we postulated that HP (2-20) might be possessed virus-cell fusion inhibitory activity. To develop the novel virus-cell fusion inhibitory peptides, several analogues with amino acid substitution were designed to increase or decrease only net hydrophobic region. In particular, substitution of Gln and Asp for hydrophobic amino acid, Trp at position 17 and 19 of HP (2-20) (Anal 3) caused a dramatic increase in virus-cell fusion inhibitory activity without hemolytic effect.
[Back to top] Structure-Antibacterial Activity Relationship
of Cecropin A Derivatives
Song Yub Shin, Keun Woo Lee, Yangmee Kim, Jae II Kim,
Kyung-Soo Hahm and Shin-Won Kang
To investigate the
effect of net positive charge, a-helicity and hydrophobicity of the peptides
on antibacterial activity, we designed three kinds of cecropin A (CA)
derivatives. Compared to CA, F3 with the highest net positive charge exhibited
similar or slightly weaker antibacterial activity (MIC: 3.13~6.25 mM). F1 showed lower antibacterial activity
than that of F3, even though it has higher hydrophobicity and a-helicity than F3. This result indicates that
the net positive charge of cecropin A-like peptides appears to be a more
important factor in antibacterial activity than a-helicity and hydrophobicity. The two
peptides F1 and F2 possessed almost similar antibacterial activity, but F2
showed very lower activity in the membrane disruption than F1, suggesting other
factors are involved in the antibacterial activity of the peptides as well as
peptide-lipid interaction.
[Back to top]
A Model for the Cytoplasmic Domain of the Influenza A
Virus M2 Channel by Analogy to the HIV-1 Vpu Protein
José W. Saldanha, Peter E. Czabotar, Alan J. Hay and
William R. Taylor
A model for the
cytoplasmic domain of the M2 proton channel of influenza A virus was formulated
based primarily on the cytoplasmic domain of the Vpu protein of HIV-1 using
sequence similarity and structure prediction techniques. The model consists of
a pair of antiparallel helices followed by a strand parallel to the first
helix. Structural analogies with other proteins contribute support for features
of the model and suggest ways in which the M2 cytoplasmic domain can interact
with other viral and cellular factors.
[Back to top] Selective Precipitation of Haptoglobin and a2-Macroglobulin from Human Serum Using
Alocasia macrorhiza Tuber Protein
B. Shivananda Nayak, N. Jagadish Ulloor and B. Shivaraj
Treatment of human
serum with ammonium sulfate fraction (0-50%) of Alocasia macrorhiza tuber
extract resulted in precipitation at neutral pH. The precipitate was dissolved at pH 10.5 and chromatographed on
Sephadex G-100 column. Two protein peaks were resolved. While the first peak
represented a2-macroglobulin and haptoglobin, the second peak accounted for specific
Alocasia protein. Incidentally the Alocasia protein was shown to be responsible
for selective and specific precipitation of a2-macroglobulin and haptoglobin from
serum. Thus the plant protein in its
pure form or in crude stage could be used for the rapid isolation of two of the
prominent a2-globulins.
[Back to top] Biochemical Characterization of Apoptotic
Cleavage of KH-Type Splicing Regulatory Protein (KSRP)/Far Upstream
Element-Binding Protein 2 (FBP2)
Heeyoung Seok, Jinsun Cho, Minseok Cheon and II-Seon
Park
Caspases,
Asp-specific cysteine protease, cleave proteins upon apoptosis. To identify and
characterize new caspase substrate in the nucleus, the proteome of the rat
liver extracts was analyzed after the treatment with caspases. One of the
identified proteins was KSRP/FBP2 that is preferentially cleaved by caspase-3
and –7 at two sites after Asp102 and Asp183. The second site was cleaved only
in the protein produced in cells, but not in in vitro translated protein. These
results indicate that more than the primary sequence may be important for the
recognition by caspases.
[Back to top] Acyl-Gelatins for Cell-Hybrid Biomaterials:
Preparation of Gelatins with High Melting Point and Affinity for Hydrophobic
Surfaces
Keiichi Miyamoto, Hiroko Chinzei and Takashi Komai
In the development
of cell-hybrid biomaterials, the functional activity of cells depends on the
selective binding of cells to artificial ligands on the biomaterials. The
extracellular matrix (ECM) is the most important ligand for cell activity. ECM
is known to contain collagen, one of whose constituents is gelatin. Although
natural gelatin has good cell attachment properties, the melting point of
gelatin hydrogel is lower than body temperature. Thus, non-chemically
cross-linked gelatin hydrogel is not a biomaterial that is used for prostheses.
In the present study, we report the preparation of acyl-gelatin hydrogels with
high melting point (>37˚C) and high affinity for hydrophobic surfaces
for easy handling for transportation and adhesion activities on the hydrophobic
surfaces. In addition, the doubling time of endothelial cells on the coated
cell culture plate was faster than that of natural gelatin owing to the higher
adhesion activity of acyl-gelatin. The results clearly demonstrated that the
acyl-gelatin acted as an interface that enabled cell adhesion to artificial
materials surfaces.
[Back to top] Synthesis of Alkyl and Aryl Esters of
N-Protected b-Homoamino Acids
from N-Protected a-Aminodiazoketones
G.
R. Vasanthakumar, H. N. Gopi and V. V. Suresh Babu
A simple and
concomitant esterification method for the synthesis of methyl, ethyl, t-butyl,
benzyl, and 9-fluorenylmethyl esters of Fmoc-/Boc-/Z-b-homoamino acids employing Fmoc-/Boc-/Z-a-aminodiazoketones by Wolff rearrangement is
described. The method offers good yield with purity.
[Back to top] Antibiotic Properties of Novel Synthetic
Temporin A Analogs and a Cecropin A-Temporin A Hybrid Peptide
David Wade, Jan-Ingmar Flock, Charlotta Edlund,
Ingegerd Löfving-Arvholm,Matti Sällberg, Tomas Bergman, Angela Silveira,
Cecille Unson,Louise Rollins-Smith, Jerzy Silberring, Malcom Richardson, Pentti
Kuusela and Hilkka Lankinen
A, 18 analogs, and
a cecropin A-temporin A hybrid peptide were tested with antibiotic sensitive
and resistant bacteria, fungi, human erythrocytes, and in clotting
assays.Several peptides were active in these assays, and some analogs (D-TA,
W1-TA, and Con-L4,G10) may be useful lead compounds for further antibiotics
development.The activity of temporin A was found to be dependent upon several
of its structural features, including amino acid composition and sequence,
chirality, helicity, and positive charge.
[Back to top] A Direct Continuous pH-Spectrophotometric
Assay for Arginine Kinase Activity
Yu Z., Pan J. and Zhou H.M.
A direct and continuous spectrophotometric method was developed for determining arginine kinase (phosphoarginine synthesis) activity. Protons are produced during the phosphoarginine synthesis course, so adding the complex acid-base indicator to this solution and monitoring the decrease of absorbance of the solution at 575 nm will follow the arginine kinase activity. For this condition, one activity unit of arginine kinase was defined as 1 mmol H+ produced in 1 minute in the enzymatic reaction catalyzed by arginine kinase.
[Back to top] Purification, Crystallization and Preliminary
X-Ray Studies of Human Augmenter of Liver Regeneration
Chao-Neng Ji, Zai-Long Cai, Gen-Tao Cao, Gang Yin,
Bing-Hua Jiao Tao Jiang, Guang Shu, Ji-Fang Mao, Yi Xie, and Yu-Min Mao
Human augmenter of
liver regeneration has been expressed in Escherichia coli, purified and
crystallized. The crystals belong to space group C222, with
unit-cell parameters a=51.7 Å, b=78.8 Å, c=63.7 Å. Diffraction data were
collected to 2.80 Å with a completeness of 99.9% (99.9% for the last shell), a
Rsym value of 0.092(0.236) and an I/s(I) value of 6.2(2.7).