Ceruloplasmin,
The Unique Multi-Copper Oxidase of Vertebrates. Pp159-169
Giovanni Musci
Predicted Fold for the ABri
Amyloid Subunit: A Model for Amyloidogenesis in Familial British Dementia.
Pp171-178
Daruka Mahadevan, Tapan Chattopadhyay, Rex A.
Palmer, Ronan O’Brien and José W. Saldanha.
Comparative
Biochemical Studies of Myotoxic Phospholipase A2 From Bothrops Venom
Pp.179-186
M.H.Toyama, A.M.Soares, S.H.Andriao-Escarso,
J.C.Novello. B. Oliveira, J.R.Gilglio, M.R.M. Fontes and S.Marangoni.
Purification
And Identification of A 25 kDa Hemorrhagin From B. atrox Venom Pp. 187-192
Jorge H.Petretski, Milton M. Kanashiro, Flavia G.
Rodriques , Elias W. Alves ,Olga L.T. Machado, and Tereza L. Kipnis
Comparative
Studies of three Type II Ribosome-inactivating Proteins from the Seeds of three
species of the Genus Cinnamomum Pp. 193-200
Fa-jian Hou, Bao-zhong Wang, Wang-yi Liu
Cloning
and Expression of a Novel Long Neurotoxin from Bungarus multicinctus in E.coli Pp.
201-207
Qin Cai, Sheng-li Yang and Yi Gong
Characteristics
of a New Antifungal Protein from Ginkgo biloba L. Pp. 209-216
Hu Huang, Jin-fei Xu and Zhen-zhen Gong
Functional
Expression of a Galactose-binding C-Type Lectin in Pichia pastoris Pp.
217-224
Jia-da Li, Qiang Xu and Ke-yi Wang
The
Performance of Callosobruchus maculatus F. In Artificial Seeds Containing
Proteins From Prosopis juliflora Seeds Pp. 225-229
Adeliana S. Oliveira, Liziane M. Lima, Railene A.
Pereira, Ana Heloneida A. Morais, Lucilene S. Pereira and Maurício P. Sales
Novel
Cross-Linking Agents for Hydrogelation of Biological Polymers Pp231-236
K.Miyamoto, M.Tokita and T.Komai
Crystallization
And Preliminary X-Ray Craystallographic Studies of Formaldehy Dehydrogenase from Pesudomonas putida Pp
237-240
Yoshio Kusakabe, Nobutada Tanaka, Kiyoshi Ito,
Tadashi Yoshimoto, and Kazuo T.Nakamura.
[Back to top] Ceruloplasmin,
The Unique Multi-Copper Oxidase of Vertebrates
Giovanni Musci
This review shortly summarizes the structural and functional properties of ceruloplasmin, the blue copper oxidase of vertebrates. Particular emphasis will be put on the unique copper stoichiometry of ceruloplasmin in comparison to all other multi-copper oxidases, and different hypotheses will be discussed to explain the apparently useless redundancy of type 1 copper ions in the vertebrate oxidase.
[Back
to top] Predicted Fold for the ABri Amyloid Subunit: A Model for
Amyloidogenesis in Familial British Dementia.
Daruka Mahadevan, Tapan Chattopadhyay, Rex A.
Palmer, Ronan O’Brien and José W. Saldanha.
ABri is a
34-residue polypeptide that forms amyloid fibrils in Familial British Dementia
(FBD). A PSI-Blast search, 3 different fold recognition programs and 3D model
building indicated that it has 3 b-strands forming an antiparallel b-sheet and a small
C-terminal a-helix.
In order to validate the prediction, the region coding the ABri polypeptide was
cloned, expressed and purified from E.Coli. Circular Dichroism (CD)
spectroscopy of ABri in aqueous solution shows it to have a predominantly b-sheet
structure with a small a-helical component.
[Back
to top] Comparative Biochemical Studies Of Myotoxic Phospholipase A2
From Bothrops Venom
M.H.Toyama, A.M.Soares, S.H.Andriao-Escarso,
J.C.Vovell. B. Oliveira, J.R.Gilglio, M.R.M.Fontes and S. Marangoni
Venoms from
Bothrops jararcussu Bothrops asper
Bothrops atrox Bothrops pirajai Bothrops moojeni Bothrops alternatus and
Bothrops (Bothriopasis) bilineata were fractionated using a simplified
procedure based on-exchange chromatography on CM-Sepharose at pH 8.0 or reverse
phase HPLC.The resulting elution profilesshowed important differences in the
myotoxin contentof these venoms.The venoms froms B.alternatus,B.atrox and
Bothriopsis bilineata did not contain the major myotoxin found in the other
venoms.The amino acid sequences of the first 50 residues of the N-terminal
region of the PLA20Like mytoxins showed a homology of 90-96% with other bothropic
myotoxins.All of the myotoxins isolated induced rat paw edema, increased the
level of plasma ceratine kinase and produced myonecrosis together with
polymorphoniclear cell infiltration.
[Back to top] Purification
And Identification Of A 25 Kda Hemorrhagin From B. atrox Venom
JorgeH.Petretski, Milton M. Kanashiro ,Flavia G.
Rodriques , Elias W. Alves , Olga L.T. Machado, and Tereza L. Kipnis
Viperine and
crotaline snake venoms contain one or more hemorrhagic metalloproteinases
called hemorrhagins. The most potent hemorrhagins belong to P-III class and
have, in adition to the protease domain, disintegrin-like and cysteine-rich
domains. Although proteolytic degradation of vascular endothelium basement
membrane has been established to be the main factor responsible for hemorrhage,
several studies reveal other factors that actually do facilitate this process.
In this study we report the identification of an P-I class hemorrhagin from
Bothrops atrox venom. We have formerly purified an P-III
class hemorrhagin from the same venom. Although exhibiting a higher proteolytic
activity, the P-I class hemorrhagin showed to be a less efficient hemorrhagin
when compared in vivo with the previously described P-III class
metalloprotease.
[Back
to top] Comparative Studies of three Type II Ribosome-inactivating
Proteins from the Seeds of three species of the Genus Cinnamomum
Fa-jian Hou, Bao-zhong Wang, Wang-yi Liu
Bodinierin, one
of the major proteins in the kernels of camphor tree (Cinnamomum Bodinieri),
has been identified as a novel type II ribosome-inactivating protein. Using
sepharose-4B column chromatography followed by acid precipitation and ammonium
sulfate precipitation, bodinierin has been purified to be homogeneous as
characterized by SDS-PAGE. Bodinierin was composed of two chains (A- and
B-chain) with the molecular weight of 31 and 34 kDa respectively. The reduced
bodinierin showed strong inhibitory activity to protein synthesis in the rabbit
reticulocyte lysate and the RNA N-glycosidase activity. The bodinierin also
displayed the carbohydrate binding activity to agglutinate rabbit erythrocyte.
A comparison of bodinierin with cinnamomin and porrectin that were isolated
from the kernels of other species of the same genus (Cinnamomum) demonstrated
that they had similar structure and biological activities, which provided
phylogenetic evidence to the three species.
[Back
to top] Cloning and Expression of a Novel Long Neurotoxin from
Bungarus multicinctus in E.coli
Qin Cai, Sheng-li Yang and Yi Gong
The total RNA was isolated from Bungarus multicinctus venom. A novel long neurotoxin (LNT) cDNA was amplified by RT-PCR method. Sequence analysis revealed that the cDNA of long neurotoxin has an open reading frame encoding 94 amino acids, including a signal peptide of 21 amino acids. The mature peptide encoding sequence was inserted into plasmid pET-28 (a+), and then which was expressed in the E. coli BL21 (DE3). The expressed protein is identified as a novel long neurotoxin by western blot.
[Back
to top] Characteristics
Of A New Antifungal Protein From Ginkgo Biloba L.
Hu Huang, Jin-fei Xu and Zhen-zhen Gong
A new antifungal
protein, named as GAFP-1, was purified from leaves of Ginkgo biloba L.. Its
molecular weight identified by mass spectrometry is 25988.0 Da and pI is 4.3.
The analysis of amino acids composition showed it has high percentages of Gly
and Glu/Gln. The 15 N-terminal amino acid residues of GAFP-1 were determined.
GAFP-1 could inhibit the hyphal growth of P. sasakii Ito, the IC50
towards P.sasakii Ito is 250µg/ml. GAFP-1 has chitinase activity, the optimal
pH and temperature being 5.0 and 50°C
[Back to
top] Functional
Expression of a Galactose-binding C-Type Lectin in Pichia pastoris
Jia-da Li, Qiang Xu and Ke-yi Wang
Trimeresurus
stejnegeri lectin (TSL) was expressed in the yeast Pichia pastoris under the
control of alcohol oxidase (AOX1) promoter. The recombinant protein, which was
glycosylated, can be purified by one-step affinity chromatography. The purified
recombinant TSL shared similar sugar-binding activity and specificity with the
native protein as determined by hemagglutination and enzyme-linked lectin
binding assays.
[Back
to top] The Performance Of Callosobruchus Maculatus F. In Artificial
Seeds Containing Proteins From Prosopis Juliflora Seeds
Adeliana S. Oliveira, Liziane M. Lima, Railene A.
Pereira, Ana Heloneida A. Morais, Lucilene S. Pereira and Maurício P. Sales
This work
reportes the effects of isolated proteins from algaroba (Prosopis juliflora
D.C.) seeds fractionated by ammonium sulfate at saturation 0-30%, 30-60% and
60-90%. They were tested, in vitro, against papain and midgut homogenate of C.
maculatus larvae. Also, the effects in vivo these protein fractions on the
development of C. maculatus larvae were tested. F 30/60 fraction was effective
against papain (72% inhibition), midgut
homogenate of larvae of C. maculatus (59,2% inhibition) and in
vivo with WD50 of
1,7%.
[Back
to top] Novel Cross-Linking Agents For Hydrogelation of Biological
Polymers
K.Miyamoto, M.Tokita and T.Komai
Cross-linking agents are useful for investigating the biological functions of biological polymers. Here we would like to report the preparation of a 4-hydroxyphenyldimethyl-sulfoniummethlysulfate (DSP) activated cross-linker that can cross-link proteins as a novel cross-linking agents. By using this cross-linker, gelatin can be cross-linked both in the sol-state at 400C and in the gel-state at 20C. The chemically cross-linked gelatin gels by DSP-activated esters that were obtained in gel-state and sol-state at a gelatin concentration greater than 1% and 5%, respectively. The fixed conformation of the folding structure seen with this physical cross-linking system is thought to result from the structure of the native protein. In addition, the urea concentration dependence of the swelling ration indicates that the hydrogen bonds are much more stable in the gel prepared from the gel state of gelatin than those in the gel prepared from the sol state of gelatin. As result, these novel bifunctional agents of the hydrogelation of biological polymers are expected application in studying intermolecular interaction.
[Back
to top] Crystallization
And Preliminary X-Ray Craystallographic Studies Of Formaldehyde Dehydrogenase
From Pesudomonas Putida
Yoshio Kusakabe, Nobutada Tanaka, Kiyoshi Ito,
Tadashi Yoshimoto, and Kazuo T.Nakamura.
The
formaldehyde dehydrogenease from Pseudomonas putida (PFDH), has been
crystallized by the vapour diffusion method using ammonium sulfate as the
Precipitating agent. The crystals belong to a trigonal space group P3112
(or P32 12) with the cell dimensions of a = b = 85.74 Ǻ,
and c = 190.9 Ǻ. There are two subunits per asymmetric unit. The
crystals diffract to at least 2.2 Ǻ resolution using Cu K a radiation at 100 K. Self-rotation function
studies suggest that the tetrameric PFDH molecular has the 222 point group
symmetry.