Protein & Peptide Letters, Volume 10, No. 5, 2003
Synthesis
of Retro Acyl Carrier Protein (74-65) Fragment on a New Glycerol Based
Polystyrene Support
Pp.427-433
P.G.
Sasikumar, K.S. Kumar and V.N. Rajasekharan Pillai
A
Sequence Function Reveals New Features in b-Protein Folding Pp.435-439
Hui
Shao and Zong-Hao Zeng
Multiple
Roles of Glutathione Binding-Site Residues of Glutathione S-Transferase Pp.441-448
Ardcharaporn
Vararattanavech and Albert J. Ketterman
A New
Dehydrogenase Specific Towards Aromatic Aldehydes from a Halophilic Bacterium Pp.449-457
F.
La Cara , L. Alves , F. Girio , A. Di Salle , A. Capasso and M. Rossi
Direct
Screening of Libraries of Yeast Clones for a-Amylase Activity on Raw Starch
Hydrolysis Pp.459-468
Dominic
W.S. Wong, Sarah B. Batt, Charles C. Lee and George H. Robertson
In
Vivo Lipoprotein Binding Assay of the Insect Exchangeable Apolipoprotein,
Apolipophorin-III Pp.469-473
Palaniappan
S. Chetty, Estela L. Arrese and Jose L. Soulages
Anticonvulsant
Activity of Benzylamides of Some Amino Acids and Heterocyclic Acids Pp.475-482
Ryszard
Paruszewski, Marzanna Strupinska, Grazyna Rostafinska-Suchar and James P.
Stables
Extracellular
Domain of Myelin Oligodendrocyte Glycoprotein (MOG) Exhibits Solvent-Dependent
Conformational Transitions
Pp.483-490
Maria
Ngu-Schwemlein, Michele Corzett, Kevin H. Thornton, Rod Balhorn and Monique Cosman
Regulation
of in Vitro Fibril Formation of Synuclein Mutant Proteins by hsp104p Pp.491-495
Byungmoon
Kong, Young Kee Chae and Kyunghee Lee
Factors
Determining the Efficacy of Alphahelical Antimicrobial Peptides Pp.497-502
Sarah
R. Dennison, Frederick Harris and David A. Phoenix
The
Role of Tyrosine Residues in the RNA N-Glycosidase Activity of Cinnamomin
A-Chain Pp.503-509
Hong Xu and Wang-Yi Liu
Crystallization
and Preliminary X-Ray Analysis of Class II Fructose-1,6-Bisphosphate Aldolase
from Thermus Caldophilus
Pp.511-515
Jun
Hyuck Lee, Young Jun Im, Seong-Hwan Rho, Seong Ho Park, Mun-Kyoung Kim, Su Jin
Cho, Tae-Yeon Kim, Jin Hwan Oh, Hyun-Jae Shin,
Crystallization
and Preliminary X-Ray Diffraction Analysis of Phosphoglucose Isomerase from
Pyrococcus Furiosus
Pp.517-520
Michael K. Swan, Thomas Hansen, Peter Schonheit and Christopher Davies
Purification,
Crystallization and Preliminary X-Ray Studies of a p- Nitrophenylphosphatase from
Bacillus Stearothermophilus
Pp.521-524
Chao-Neng Ji, Liang Tian, Cong-Jing Feng, Gang Yin, Guang Shu, Ji-Xi Li, Wei-Ming Gong, Hai Pang, Yi Xie and Yu-Min Mao
Initiating
Structural Studies of Lys49-PLA2 Homologues Complexed with an Anionic
Detergent, a Fatty Acid and a Natural Lipid Pp.525-530
L. Watanabe , M.R.M. Fontes , A.M. Soares , J.R. Giglio and R.K. Arni
[Back to top] Synthesis of Retro Acyl Carrier
Protein (74-65) Fragment on a New Glycerol Based Polystyrene Support Pp.427-433
P.G. Sasikumar, K.S. Kumar and V.N. Rajasekharan Pillai
Retro-ACP (74-65)
fragment was synthesized on a novel 4% tri-(propylene glycol) glycerolate
diacrylate cross-linked polystyrene (PS-TRPGGDA) support. The peptide is grown
from the functional site present in the cross-linker, which makes it unique and
cost-effective among other styrene based polymer supports. A comparative study
with Merrifield resin indicates high yield and purity of the peptide
synthesized on the novel support.
[Back to top] A Sequence Function Reveals New
Features in b-Protein Folding
Hui
Shao and Zong-Hao Zeng
When amino acid
residues are represented by parameters describing their side chain lengths and
polarities, a sequence function defined as the sum of the first two sequence
autocorrelation functions is found to be negatively and linearly correlated
with the logarithms of folding rates of b-proteins. The new function reveals
new features in b-protein folding: larger residues slow down the folding while
alternative distribution of polar-non-polar residues accelerates the folding.
[Back to top]
Multiple Roles of
Glutathione Binding-Site Residues of Glutathione S-Transferase
Ardcharaporn Vararattanavech and Albert J. Ketterman
This study was
designed to characterize residues in the glutathione binding site of AdGSTD4-4
from the mosquito malaria vector Anopheles dirus. The data revealed that Leu33,
His38 and His50 each play a role in enzyme catalysis and glutathione binding.
The mutants of these three residues also displayed differences in hydrophobic
substrate specificity, suggesting that changes in the active site conformation
occurred. Differences in conformations was also suggested by protein stability
changes. These results indicate that residues in the glutathione binding site are
not only important in the catalytic function but also play a role in the
structural integrity of the enzyme.
[Back to top]
A New Dehydrogenase
Specific Towards Aromatic Aldehydes from a Halophilic Bacterium
F.
La Cara , L. Alves , F. Girio , A. Di Salle , A. Capasso and M. Rossi
A new enzyme
showing a dehydrogenase activity towards aromatic aldehydes was isolated,
purified and characterized from a halophilic strain isolated from saline
environment. The enzyme is a monomer of 54 kDa; it is rather thermostable
(optimal temperature: 50°C) showing a broad spectrum of activity in a large pH
range with the maximum at pH 9.5. The substrate specificity and the effect of
ions were evaluated and compared with analogous described proteins.
[Back to top] Direct Screening of Libraries of Yeast Clones for a-Amylase Activity on Raw Starch
Hydrolysis
Dominic
W.S. Wong, Sarah B. Batt, Charles C. Lee and George H. Robertson
High-throughput
screening for high-activity barley a-amylase mutants expressed in Saccharomyces
cerevisiae is hampered by the interference of reducing agents, particularly the
glucose used in yeast growth media. The present investigation employed
colorimetric and chemiluminescent detection systems that enable direct and
rapid screening of activities on raw starch substrate. Active clones could be
separated into two groups, based on high total activity or high specific
activity.
[Back
to top] In Vivo Lipoprotein Binding Assay
of the Insect Exchangeable Apolipoprotein, Apolipophorin-III
Palaniappan
S. Chetty, Estela L. Arrese and Jose L. Soulages
An original method
for the study of the lipid binding properties of exchangeable apolipoproteins
is reported. Binding of Locusta migratoria apolipophorin-III to Manduca sexta
low-density lipophorin (LDLp) and high-density lipophorin (HDLp) was studied in
vivo. This assay could be used useful to investigate the effect of mutations in
the lipid binding properties of exchangeable apolipoproteins under
physiological conditions.
[Back to top] Anticonvulsant Activity of
Benzylamides of Some Amino Acids and Heterocyclic Acids
Ryszard
Paruszewski, Marzanna Strupinska, Grazyna Rostafinska-Suchar and James P.
Stables
A series of new
potential anticonvulsants have been synthesized. They are N-methyl benzylamides
of N-methyl Asp and N-methyl Glu (R and S), benzylamides of some heterocyclic
acids and their Noxides and benzylamides of two heteroalicyclic acids. The
obtained compounds were evaluated in the Anticonvulsant Screening Project (ASP)
of Antiepileptic Drug Development Program (ADDP) of NIH.
[Back to top] Extracellular Domain of Myelin
Oligodendrocyte Glycoprotein (MOG) Exhibits Solvent-Dependent Conformational
Transitions
Maria
Ngu-Schwemlein, Michele Corzett, Kevin H. Thornton, Rod Balhorn and Monique Cosman
The conformation
of the non-glycosylated recombinant form of the extracellar domain of rat MOG
(rMOG(1-125)) dissolved in different solvent conditions was studied by CD
spectroscopy. The results show that rMOG(1-125) exhibits a predominantly b
sheet conformation in aqueous buffer solution at pH 7.5 and that this 'b-form'
is stabilized by zwitterionic phospholipids, DPC and LPCP. The a helical
content of the protein can increase from 9% to up to 20% when TFE or anionic
detergent LPAP and SDS are added.
[Back to top] Regulation of in Vitro Fibril
Formation of Synuclein Mutant Proteins by hsp104p
Byungmoon
Kong, Young Kee Chae and Kyunghee Lee
Hsp104p, as an
anti-oxidative protector of ROS generation, was examined to inquire if it
prevents aggregation of synuclein mutants, A30P or A53T upon aging, in vitro.
The role of Hsp104p was also addressed in dissociation of pre-formed aggregates
of synuclein mutants. Significant protection in fibril formation was observed by
wild-type Hsp104p regardless of ATP presence, not by mutant Hsp104p. To a
lesser extent, the dissociation effect of wild-type Hsp104p was observed only
in the presence of ATP. These results will be discussed in relation to the
development of an antioxidant approach to prevent amyloid fibril formation in
several neurodegenerative diseases.
[Back to top] Factors Determining the Efficacy
of Alphahelical Antimicrobial Peptides
Sarah
R. Dennison, Frederick Harris and David A. Phoenix
A database of a-helical antimicrobial peptides (AMP) was established and their minimum
inhibitory concentrations (MIC) were compared with their physiochemical
characteristics in an attempt to establish those features that determine
efficacy. There is no significant difference in AMP sensitivity between
Gram-positive and Gram-negative bacteria but fungi did require higher
concentrations to achieve the same degree of growth inhibition. For
antibacterial peptides there appears to be a positive correlation between MIC
and hydrophobic arc size and a negative correlation between MIC and net charge.
[Back to top] The
Role of Tyrosine Residues in the RNA N-Glycosidase Activity of Cinnamomin
A-Chain
Hong
Xu and Wang-Yi Liu
Cinnamomin is a
type II ribosome-inactivating protein (RIP) and its A-chain (CTA) is a RNA
Nglycosidase. It is observed that modification of tyrosine residues by
N-acetylimidazole (N-AI) causes almost complete loss of CTA activity. Adenine
partially protects tyrosine residues from modification by NAI. It is proposed
that tyrosine residues are involved in the active site of CTA and they are
crucial in recognition and binding of ribosomal RNA. Tryptophan residues of CTA
are also studied by NBS modification.
[Back to top] Crystallization and Preliminary
X-Ray Analysis of Class II Fructose-1,6-Bisphosphate Aldolase from Thermus
Caldophilus
Jun
Hyuck Lee, Young Jun Im, Seong-Hwan Rho, Seong Ho Park, Mun-Kyoung Kim, Su Jin
Cho, Tae-Yeon Kim, Jin Hwan Oh, Hyun-Jae Shin,
In this study, we
have crystallized class II fructose-1,6-bisphosphate aldolase (FBA) from
Thermus caldophilus (Tca). Purified Tca FBA is a tetrameric enzyme of 305
residues, which crystallizes in the space group P212121 (cell dimensions a =
98.9, b = 113.1, c = 115.7 Å), with four molecules in the asymmetric unit. A
complete diffraction data set was obtained from orthorhombic crystals at
resolution of 2.2 Å.
[Back to top] Crystallization
and Preliminary X-Ray Diffraction Analysis of Phosphoglucose Isomerase from
Pyrococcus Furiosus
Michael
K. Swan, Thomas Hansen, Peter Schonheit
and Christopher Davies
In several
euryarchaeota, phosphoglucose isomerase (PGI) activity is catalyzed by an
enzyme unrelated to the well-known family of PGI enzymes found in prokaryotes,
eukaryotes and some archaea. In order to understand the mechanistic differences
between the two families of enzymes we have crystallized PGI from the archaeon
Pyrococcus furiosus. The crystals belong to the space group P21 and a complete
dataset extending to 1.9 Å resolution has been collected.
[Back to top] Purification,
Crystallization and Preliminary X-Ray Studies of a p- Nitrophenylphosphatase
from Bacillus Stearothermophilus
Chao-Neng
Ji, Liang Tian, Cong-Jing Feng, Gang Yin, Guang Shu, Ji-Xi Li, Wei-Ming Gong,
Hai Pang, Yi Xie and Yu-Min Mao
Thermostable p-nitrophenylphosphatase from Bacillus stearothermophilus has been expressed in
Escherichia coli, purified and crystallized. The crystals belong to space group
C2, with unit-cell parameters a = 67.17 Å, b = 57.84 Å, c = 62.49 Å and
a =
90.0º, b = 95.4º, g = 90.0º. Diffraction data were collected to 1.40 Å
resolution with a completeness of 94.7% (96.6% for the last shell), an Rfac
value of 0.074 (0.341) and an I/s (I) value of 30.1 (2.67).
[Back to top] Initiating Structural Studies of
Lys49-PLA2 Homologues Complexed with an Anionic Detergent, a Fatty Acid and a
Natural Lipid
L. Watanabe , M.R.M. Fontes , A.M. Soares , J.R.
Giglio and R.K. Arni
Lys49-Phospholipase
A2 (Lys49-PLA2 - EC 3.1.1.4) homologues damage membranes by a Ca2+- independent
mechanism which does not involve catalytic activity. Both MjTX-II from Bothrops
moojeni and BthTX-I from Bothrops jararacussu are dimeric in solution and in
the crystalline states, and a model for the Ca2+-independent membrane damaging
mechanism has been suggested in which flexibility at the dimer interface region
permits quaternary structural transitions between “open” and “closed” membrane
bound dimer conformations which results in the perturbation of membrane phospholipids
and disruption of the bilayer structure [1]. With the aim of gaining insights
into the structural determinants involved in protein/lipid association, we
report here the crystallization and preliminary X-ray analysis of the (i)
MjTXII/ SDS complex at a resolution of 2.78Å, (ii ) MjTX-II/STE complex at a
resolution of 1.8 Å and (iii) BthTXI/ DMPC complex at 2.72Å. These complexes
were crystallized by the hanging drop vapour-diffusion technique in (i) HEPES
buffer (pH 7.5) 1.8M ammonium sulfate with 2% (w/v) polyethyleneglycol 400, in
(ii ) 0.6-0.8 M sodium citrate as the precipitant (pH 6.0-6.5) and in (iii)
sodium citrate buffer (pH 5.8) and PEG 4000 and 20% isopropanol, respectively.
Single crystals of these complexes have been obtained and Xray diffraction data
have been collected at room temperature using a R-AXIS IV imaging plate system
and graphite monochromated Cu Ka X-ray radiation generated by a Rigaku RU300
rotating anode generator for (i) and (iii) and using using a Synchrotron
Radiation Source (Laboratório Nacional de Luz Sincrotron, LNLS, Campinas,
Brazil) for (ii ).