An Alternative Method for the Isolation and Identification of C-Terminal
Peptide of Protein. Pp. 195-202.
Kuo-Liang His, Lydia M. Nuwaysir and David R. Dupont
[Abstract]
Carboxyl-Terminal Sequencing: COOH-Amino Acid Activation with DCC or
TBTU. Pp. 203-208.
Patricia D. Ribeiro, Elias W Alves and Olga L.T Machado
[Abstract]
Synthesis of Two Designed Hairprin Peptides on a Newly Developed 1,6-Hexanediol
Diacrylate (HDODA)-Crosslinked Polystyrene Resin. Pp. 209-214.
Jaya F. Varkey, Satish Kumar Awasthi and V.N. Rajasekharan Pillai
[Abstract]
Interaction of the Seed Lectin from Parkia platycephala (Mimosoideae)
with Carbohydrates and Complex Glycans. Pp. 215-222.
Márcio V Ramos, Benildo S. Cavada, Liezelotte R. Bomfim,
Henri Debray, Anne-Marie Mazard, Juan J. Calvete, Thalles B. Grangeiro
and Pierre Rougé
[Abstract]
Purification and Partial Characterization of 5-Oxo-L-Prolinase from
Pig Kidney. Pp. 223-228.
Martina Jäger, Peter Weber, Thorsten Bangsow and Sabine Wolf
[Abstract]
Binding of l-Anilino-8-Napthalenesulfonate to Free and Coformycin-Complexed
Bovine Adenosine Deaminase. Pp. 229-232.
Brain Cannon, Jesse A. Oakes and B. Mark Britt
[Abstract]
Zinc Promoted Simple Synthesis of Z-Amino Acids Under Neutral Conditions.
Pp. 233-236.
Hosahudya N. Gopi, Kuppanna Ananda and Vommina V Suresh Babu
[Abstract]
The a-Fetoprotein Molecule has One or Two
Rigid Domains Depending on the Protein Purification Procedure. Pp. 237-244.
Andrey Yu. Tomashevski, Tatyana N. Melnik, Natalya V. Narizhneva,
Mikhail M. Shavlovsky, Vadim B. Vasiliev and Vladimir N. Uversky
[Abstract]
Design, Synthesis and DNA-Binding Activities of GCN4 Peptide Dimer Studied
by Circular Dichroism Spectroscopy. Pp. 245-252.
He Wang, Luhua Lai, Zhenwei Miao and Xiaojie Xu
[Abstract]
Crystallisation Report
Purification and Crystallisation of a Novel Two-Domain Lectin from Scilla
Campanulata. Pp. 253-258.
Lisa M. Wright, Cohn D. Reynolds, Pierre J. Rizkallah, Anthony K.
Allen, Willy J. Peumans, Els Van Damme and Michael J. Donovan
[Abstract]
[Back to top] An Alternative
Method for the Isolation and Identification of C-Terminal Peptide of Protein.
Kuo-Liang His, Lydia M. Nuwaysir and David R. Dupont.
A chemical approach for selectively isolating the C-terminal peptide
from a protein is described. The protein to be analyzed is derivatized
with glycinamide to protect carboxyl groups at both C-terminus and Asp/Glu
side chains and subsequently fragmented with protease. The resulting peptides
are treated with a Sepharose-based resin, EAH Sepharose 4B which binds
all fragments except the C-terminal peptide, through the formation of amide
bonds. The unbound C-terminal peptide is purified by HPLC and identified
using Edman sequencing and mass spectrometry analyses.
[Back to top] Carboxyl-Terminal
Sequencing: COOH-Amino Acid Activation with DCC or TBTU. Patricia D. Ribeiro,
Elias W Alves and Olga L.T Machado.
The present study explores conditions to activate the carboxyl group
before the coupling of C-terminal amino acid with Trimethylsilyl -isothiocyanate
(TMS-ITC). Protected amino acids and different peptides were used as model
and l,3'-Dicyclohexylcarbodiimide (DCC) plus 1-Hydroxybenzotriazole (HOBt)
and 2-(1H-Benzotriazol-l-y)-1,1,3,3-tetramethyluronium tetrafluoroborate
(TBTU) plus HOBt were used as carboxyl activators. The coupling reaction
of activated AA with TMS-ITC was temperature dependent and the best yield
(100 % of coupling) was obtained at 70°C, when DCC or TBTU both with
HOBt were used as activating reagents.
[Back to top] Synthesis
of Two Designed Hairprin Peptides on a Newly Developed 1,6-Hexanediol Diacrylate
(HDODA)-Crosslinked Polystyrene Resin. Jaya F. Varkey, Satish Kumar Awasthi
and V.N. Rajasekharan Pillai.
Synthesis of two designed hairpin peptides on 1,6-hexanediol diacrylate
crosslinked polystyrene support using the standard solid phase methodology
is described. Both the peptides are obtained in high yield and purity.
The new polymeric system is an ideal support for the synthesis of hairpin
peptides, which is a very difficult task by the solid phase method.
[Back to top] Interaction
of the Seed Lectin from Parkia platycephala (Mimosoideae) with Carbohydrates
and Complex Glycans. Márcio V Ramos, Benildo S. Cavada, Liezelotte
R. Bomfim, Henri Debray, Anne-Marie Mazard, Juan J. Calvete, Thalles B.
Grangeiro and Pierre Rougé.
The carbohydrate-binding speciticity of the Parkia platycephala
(Mimosoideae) lectin was investigated by different techniques including
hapten inhibition of haemagglutination, binding of lectin to immobilized
glycoproteins and interaction in real time performed by surface plasmon
resonance. The lectin belongs to the mannose/glucose-specific group and
mannose and oligomannosides with a1-3 and a1-6
linkages are among its best inhibitors. The mannose/glucose-binding activity
exhibited by this lectin, which belongs to a primitive group of Leguminosae,
could be related to a defensive function against plant pathogens.
[Back to top] Purification
and Partial Characterization of 5-Oxo-L-Prolinase from Pig Kidney. Martina
Jäger, Peter Weber, Thorsten Bangsow and Sabine Wolf.
5-Oxo-L-prolinase (E.C. 3.5.2.9) was purified to homogeneity from porcine
kidney. The molecular weight of one subunit was estimated to 135 kDa by
SDS-PAGE. N-terminal sequencing was not successful, leading to the conclusion
that the enzyme is N-terminal blocked. Partial sequence data from a chymotryptic
peptide showed strong similarities to the rat enzyme.
[Back to top] Binding
of l-Anilino-8-Napthalenesulfonate to Free and Coformycin-Complexed Bovine
Adenosine Deaminase. Brain Cannon, Jesse A. Oakes and B. Mark Britt.
We have determined the number of binding sites and average binding
affinity of the nonpolar probe 1-anilino-8-napthalenesulfonate to free
bovine adenosine deaminase and to its complex with coformycin, a potent
transition state analogue. Our results indicate an increase in average
binding affinity accompanied by a reduction in number of sites for the
coformycin-complexed enzyme. Interpretation of these results in terms of
the relative stabilities of the two enzyme states and to the catalytic
efficiency of enzymes is presented.
[Back to top] Zinc Promoted
Simple Synthesis of Z-Amino Acids Under Neutral Conditions. Hosahudya N.
Gopi, Kuppanna Ananda and Vommina V Suresh Babu.
The introduction of benzyloxylcarbonyl (Z) group into amino acids is
described at neutral pH using benzyloxylcarbonyl chloride and activated
zinc powder. The reaction is simple, fast and clean. Thus the Z-amino acids
are prepared in good yield and purity.
[Back to top] The a-Fetoprotein
Molecule has One or Two Rigid Domains Depending on the Protein Purification
Procedure. Andrey Yu. Tomashevski, Tatyana N. Melnik, Natalya V. Narizhneva,
Mikhail M. Shavlovsky, Vadim B. Vasiliev and Vladimir N. Uversky.
a-Fetoprotein samples isolated from human
cord serum by different methods were compared using circular dichroism,
scanning microcalorimetry and fluorescence spectroscopy. Their structural
properties and conformational stability were appreciably discriminated.
In particular, the protein molecule had either one or two relatively rigid
domains which can be co-operatively disrupted by heating or urea treatment.
This is due to the difference of the isolation procedures in their capabilities
to strip the natural ligands from the protein molecule.
[Back to top] Design,
Synthesis and DNA-Binding Activities of GCN4 Peptide Dimer Studied by Circular
Dichroism Spectroscopy. He Wang, Luhua Lai, Zhenwei Miao and Xiaojie Xu.
Basic region-leucine zipper (bZIP) family of DNA-binding protein GCN4
truncate Ac-[L250] GCN4 (226--252)-GGC-OH and its two designed
analogues were synthesized by solid phase method using Fmoc-strategy. Circular
dichroism spectroscopy measurements showed that the DNA-binding affinity
with target AP-1 site was reduced when Thr-236 in GCN4 basic region was
replaced by valine or glutamine.
[Back to top] Purification
and Crystallisation of a Novel Two-Domain Lectin from Scilla Campanulata.
Lisa M. Wright, Cohn D. Reynolds, Pierre J. Rizkallah, Anthony K. Allen,
Willy J. Peumans, Els Van Damme and Michael J. Donovan.
In the last decade a number of single-domain mannose-binding lectins
(11-14 kDa) have been characterized from different monocot plants and shown
to belong to a superfamily of evolutionary related proteins. We have isolated
and crystallized a novel two-domain, multivalent lectin, with 26.2 kDa
subunits, from Scilla campanulata bulbs. The native agglutinin crystallizes
in space group C2 and a 3.3Å resolution data set has been recorded.