Membrane Binding of Escherichia coli Penicillin - Binding Protein
4 is Predominantly Electrostatic and Occurs at a Specific Binding Site.
Pp. 63-66.
Frederick Harris and David A. Phoenix
[Abstract]
Effective Renaturation of MET-1 Lysozyme Expressed in Escherichia
coli as Inclusion Bodies. Pp. 67-74.
Yoshitake Maeda, Tadashi Ueda and Taiji Imoto
[Abstract]
Modeling of the Non-Obese Diabetic Mouse Class II MHC Molecule I-Ag7.
Pp. 75-82.
Wilson S. Meng, Avi V. Bhavaraju, Ian S. Haworth and Hermann von
Grafenstein
[Abstract]
Wound Healing Activity of Peptides Related to Growth Hormone Releasing
Hexapeptide. Pp. 83-86.
Bijoy Kundu, Geeta Singh, Girish K. Jain, Arti Shukla, Nidhi Shrivastava
and Gyanendra K. Patnaik
[Abstract]
The Surphagon Hydrolysis by Pepsin and Gastricsin. Pp. 87-94.
Olga Mirgorodskaya, Nataliya Savelieva, Alexandra Simankova, Andrey
Shevchenko and Sergey Alexandrov
[Abstract]
Expression and Purification of Human Metallothionein-III/Growth Inhibitory
Factor in E. coli. Pp. 95-100.
Daqing Wu, Jiong Shen and Binggen Ru
[Abstract]
Structure of Three Ribonuclease-A Covalent Derivatives with Pyridoxal
5'-Phosphate. Pp. 101-108.
Lluís Boqué, Maria G. Coll, Marc Ribó, Claudi
M. Cuchillo, Ignacio Fita and Maria Vilanova
[Abstract]
Molecular Mechanics Studies on Conformationally restricted b-Alanine
Analogs. Pp. 109-116.
Shashidhar N. Rao, Salvatore Profeta, Jr. and VN. Balaji
[Abstract]
Crystallisation Reports
Crystallization and Preliminary X-ray Analysis of Parkia pendula
Lectin. Pp. 117-120.
F.R. Lombardi, M.R.M. Fontes, G.M.O. Souza, L.C.B.B. Coelho, R.K.
Arni and WF. de Azevedo, Jr.
[Abstract]
Crystallization and Preliminary X-ray Diffraction Analysis of a LY549-PLA2
Homologue from Cerrophidion godmani Venom. Pp. 121-126.
W.F. de Azevedo, Jr., R.J. Ward, J.M. Gutiérrez, C. Díaz-Oreiro
and R.K. Arni
[Abstract]
[Back to top] Membrane
Binding of Escherichia coli Penicillin - Binding Protein 4 is Predominantly
Electrostatic and Occurs at a Specific Binding Site. Frederick Harris and
David A. Phoenix.
In a wild tvpe E. coli, MRE6OO, PBP4 is legitimately membrane
bound yet in a PBP4 over-expressor HB1O1/pBK4, less than 5% fractionates
with the membrane. It is therefore suggested that PBP4 - membrane binding
occurs at a specific binding site. Biochemical wash procedures imply that
PBP4 membrane interaction occurs via predominantly electrostatic
forces. Use of an E. coli mutant in which levels of anionic lipid
can be controlled could provide no evidence for an anionic lipid requirement.
[Back to top] Effective
Renaturation of MET-1 Lysozyme Expressed in Escherichia coli
as Inclusion Bodies. Yoshitake Maeda, Tadashi Ueda and Taiji Imoto.
The effective renaturation method that we developed was applied to
the renaturation of the reduced Met-1 lysozyme expressed in
E. coli as inclusion bodies. Using a slow dialysis renaturation
method, a concentration as high as 1.25 mg/ml of the reduced Met-1
lysozyme could be renatured in 40% yield, while the folding yield was less
than 5% even at a concentration of 0.25 mg/ml using a rapid dilution method.
[Back to top] Modeling
of the Non-Obese Diabetic Mouse Class II MHC Molecule I-Ag7.
Wilson S. Meng, Avi V. Bhavaraju, Ian S. Haworth and Hermann von Grafenstein.
A three-dimensional model of the NOD class II MHC molecule I-Ag7
was constructed based on a primary sequence alignment with HLA-DR1, another
class II MHC molecule, for which an x-ray structure has been solved. This
model was then used in the characterization of interactions of I-Ag7
with a peptide derived from mouse serum albumin (residues 560-574). The
approximate orientations of the peptide side chains were computed using
a simulated annealing approach and found to be largely consistent with
experimental data.
[Back to top] Wound Healing
Activity of Peptides Related to Growth Hormone Releasing Hexapeptide. Bijoy
Kundu, Geeta Singh, Girish K. Jain, Arti Shukla, Nidhi Shrivastava and
Gyanendra K. Patnaik.
Seven novel hexapeptides related to growth hormone releasing hexapeptide
(GHRP-6) with modifications at position 1, 3, 4 and 6 have been synthesised
and evaluated for their wound healing activity. One of the compounds (CDRI
96/257) exhibited 70% increase in the tensile strength over that of control.
[Back to top] The Surphagon
Hydrolysis by Pepsin and Gastricsin. Olga Mirgorodskaya, Nataliya Savelieva,
Alexandra Simankova, Andrey Shevchenko and Sergey Alexandro.
The comparative specificity of pepsin A and gastricsin in relation
to hydrolysis of surphagon and of some luliberin synthetic analogs containing
the D-amino acid residues was studied. According to chromatographic
and mass-spectrometric data gastricsin rather than pepsin A hydrolyzed
surphagon by the bond formed by an amino group of the DAla-residue.
The results promote to suggest that gastricsin is a highly specific proteinase
hydrolyzing structurally abnormal oligopeptides.
[Back to top] Expression
and Purification of Human Metallothionein-III/Growth Inhibitory Factor
in E. coli. Daqing Wu, Jiong Shen and Binggen Ru.
A synthetic DNA coding for human metallothionein-III (hMT-Ill)/growth
inhibitory factor(GIF) was designed for efficient expression in Escherichia
coli and cloned into the vector pGEX-4T-1. Upon induction by IPTG,
the gene for hMT-III was expressed as a glutathione-S-transferase fusion
protein which accounted for approximately 46% of the total protein in the
cells. The fusion protein was purified from both the cytosol and the inclusion
body fractions by glutathione-agarose affinity chromatography, then it
was cleaved with bovine thrombin releasing a C-terminal truncated fragment
of hMT-III. The recombinant MT-III had a high affinity for metals, and
could inhibit the growth of neuronal cells in vitro.
[Back to top] Structure
of Three Ribonuclease-A Covalent Derivatives with Pyridoxal 5'-Phosphate.
Lluís Boqué, Maria G. Coll, Marc Ribó, Claudi M. Cuchillo,
Ignacio Fita and Maria Vilanova.
The crystal structure of three bovine pancreatic ribonuclease derivatives
modified covalently with pyridoxal 5'-phosphate at Lys-1 (derivative A),
Lys-7 (derivative B) and Lys-41 (derivative C) has been determined. The
structures found provide an explanation of the kinetic pararneters obtained
for derivatives A and B. In addition, these structures permit to postulate
the contribution of Lys-7 and Lys-66 to the catalytic mechanism of ribonuclease
A.
[Back to top] Molecular
Mechanics Studies on Conformationally restricted b-Alanine
Analogs. Shashidhar N. Rao, Salvatore Profeta, Jr. and VN. Balaji.
We present molecular mechanics conformational energy calculations on
the model peptides with 2-aminocyclopropane-l-carboxylic acid (1) and 2-arninocyclobutane-1-carboxylic
acid (2). The essential results of the calculations have been presented
as plots drawn as a function of the backbone torsion angles j
and y. The low energy structures of these
model compounds provide useful insights into the design of peptidomimetics
with turn characteristics.
[Back to top] Crystallization
and Preliminary X-ray Analysis of Parkia pendula Lectin. F.R. Lombardi,
M.R.M. Fontes, G.M.O. Souza, L.C.B.B. Coelho, R.K. Arni and WF. de Azevedo,
Jr.
Lectins are proteins important in various biological processes such
as infection, cell differentiation and metastasis. The Parkia pendula
lectin has been crystallized using the hanging-drop vapour diffusion method.
X-ray diffraction data were collected using a Rigaku RU3OO rotating anode
generator and R-AXIS IV diffractometer. The cell parameters for P. pendula
lectin are a=93.7Å, b=161.1Å, c=80.0Å and space group
C222. The maximum resolution was of 2.98Å. These data showed a Rsym=l2.8%.
[Back to top] Crystallization
and Preliminary X-ray Diffraction Analysis of a LY549-PLA2 Homologue
from Cerrophidion godmani Venom. W.F. de Azevedo, Jr., R.J. Ward, J.M.
Gutiérrez, C. Díaz-Oreiro and R.K. Arni.
Lys49-Phospholipase A2 (Lys49-PLA2) homologues damage membranes
by a Ca2+-independent mechanism
which does not involve catalytic activity. The myotoxic Lys-49 phospholipase
myotoxin II from Cerrophidion
(Bothrops) godmani has been crystallized, and X-ray diffraction
data were collected to 2.8 Å resolution.
Preliminary analysis reveals the presence of one molecule in the asymmetric
unit.