ISOLATION AND CHARACTERIZATION OF TWO DISTINCT ATP-INDEPENDENT TOPOISOMERASE II ACTIVITIES FROM THE TRYPANOSOMATID TRYPANOSOMA BRUCEI BRUCEI Pp. 1-11
CHARACTERIZATION OF A HEMAGGLUTINATING GLYCOPROTEIN ISOLATED FROM BOTHROPS MOOJENI SNAKE VENOM Pp. 13-20
Bothrops moojeni VENOM PEPTIDES CONTAINING BRADYKININ POTENTIATING PEPTIDES SEQUENCE Pp. 21-26
EXPRESSION OF THE N- TERMINAL
SEGMENT OF QBRN-2 IN E.
COLI AND TIPS ON PREPARATION
OF A RECOMBINANT PROTEIN Pp. 27-32
STABILITY OF A BLACK EYED PEA TRYPSIN/CHYMOTRYPSIN INHIBITOR (BTCI) Pp. 33-38
STRUCTURAL INSIGHT INTO OXYBENZOTRIAZOLE BASED COUPLING REAGENTS. Pp. 39-44
T. K. Srivastava, W. Haq, S.
Bhanumati, D. Velmurugan, U.Sharma, N. R. Jagannathan and S. B. Katti
SIMPLE AND EFFICIENT METHOD FOR SYNTHESIS OF Z-/Boc-AMINO ACID AMIDES USING p-TOLUENESULPHONYL CHLORIDE Pp. 45-48
CD CONFORMATIONAL AND MODELING STUDIES OF A SYNTHETIC PEPTIDE VPDLLADLLK IN DIFFERENT MEDIA Pp . 49-55
CRYSTALLIZATION AND PRELIMINARY X-RAY STUDIES OF L-(+)-2,3-BUTANEDIOL DEHYDROGENASE FROM BREVIBACTERIUM SACCHAROLYTICUMC-I012 Pp. 57-61.
ULTRA-HIGH RESOLUTION X-RA Y DIFFRACTION FROM CRYSTALS OF THE KINETIC MUTANT OF HUMAN CARBONIC ANHYDRASE II, HIS 64 ALA, AND ITS COMPLEXES WITH PROTON ACCEPTOR/DONORS. Pp. 63-67.
CRYSTALLIZATION AND PRELIMINARY X-RAY DIFFRACTION STUDY OF RIBOFLA VIN SYNTHASE Pp. 69-73
[Back to top] ISOLATION AND CHARACTERIZATION OF TWO DISTINCT ATP-INDEPENDENT TOPOISOMERASE II ACTIVITIES FROM THE TRYPANOSOMATID TRYPANOSOMA BRUCEI BRUCEI
Vasanthi
R. Vittar, Anuradha Sekher, and Barbara J. Carter
In a rapid purification
procedure, two distinct type II topoisomerase activities (decatenation and
catenation) have been isolated from the
whole cell extracts of Trypanosoma brucei brucei. Both decatenating and
catenating activities were detected in
the cell extracts, but these two activities were separated in the last step of
purification. These two activities were
found to be independent of each other and competent both in the presence and
absence of I mm A TP . Key words:
catenation, decatenation, kDNA, topoisomerase, trypanosomes
[Back to
top] CHARACTERIZATION OF A
HEMAGGLUTINATING GLYCOPROTEIN ISOLATED FROM BOTHROPS MOOJENI SNAKE VENOM
B.
H. Kassab, D. D. de Carvalho, S. Marangoni and J. C. Novello
From the crude snake venom of
Bothrops moojeni, a homodimeric lectin was purified by affinity chromatography in lactose and was named BMooL. This
lectin appeared to be a glycoprotein because
it reacted with Schiff's reagent. The hemagglutinating activity of BMooL is
inhibited by galactoside, EDT A. EGT A
and DTT .The amino acid analysis of BMooL showed a high content of acidic residues and its N-terminal sequence showed
similarity with other lectins from snake venoms.
[Back to top] Bothrops moojeni VENOM PEPTIDES CONTAINING BRADYKININ POTENTIATING PEPTIDES SEQUENCE .
S.C.
Bourguignon, F.S. Pase, Jr., E.B. Calheiros,
M. Juliano., A.S. Aguiar, A.R.
Melgarejo & S. Giovanni-De-Simone
Eight peptides were isolated from
Bothrops moojeni venom by filtration followed by HPLC. MALDI mass spectroscopy and Edman degradation
determined the purity , Mr and sequence of the peptides, respectively. Three (BmvP7, BmvP8 and BmvP 11) of them were
honiologues with the biologically active
B. jararaca bradykinin potentiating peptides while the other with peptides or
protein from different sources.
[Back to
top] EXPRESSION OF THE N-
TERMINAL SEGMENT OF QBRN-2 IN E. COLI AND TIPS ON PREP ARA TION OF A
RECOMBINANT PROTEIN
Wei
Liu , Rongqiao He and Zhigang Xue.
POU box gene qBrn-2 exhibits
restricted expression pattern in quail neurogenesis and myogenesis. Here we report isolation of a N-terminal segment
(P207) of qBrn-2 expressed in E.coli and tips on preparation of a recombinant protein. Recombinant P207 was
purified and migrated as a single band on SDS-PAGE with the same apparent molecular mass as predicted. In the
expression of P207 by pET 3b vector, IPTG concentration for induction greatly affected the expression
level. Further, fusion domain at N-terminus seemed to change the stability of P207. Those results suggest some useful
tips on preparation of a recombinant protein.
[Back to top] STABILITY OF A BLACK EYED PEA TRYPSIN/CHYMOTRYPSIN INHIBITOR (BTCI)
Luciano
Paulino da Silval, Jose Roberto S. A. Leite., Carlos Bloch Jr . and Sonia Maria
de Freitas,
The stability of BTCI has been
investigated as function of pH and temperature, following its inhibitory activity against trypsin. The
isolated inhibitor of 9,084 Da is stable over pH 3 to 10 at 25°C. BTCI showed high thermal stability ranging from 25
to 95°C at pH 3.0 and 7.0. However, the protein lost about 20% of its inhibitory activity over 75°C at pH
8.2. The results indicated that BTCI is extremely stable to heat and Ph as typical ofBowman-Birk inhibitors.
[Back to top] STRUCTURAL INSIGHT INTO OXYBENZOTRIAZOLE BASED COUPLING REAGENTS.
T.
K. Srivastava, W. Haq, S. Bhanumati, D. Velmurugan, U.Sharma, N. R.
Jagannathan and S. B. Katti
Present work encompasses a detailed solution and solid phase NMR spectroscopic studies on oxybenzotriazole based coupling reagents namely HBTU (1), NBTU (2) and BOP (3). The present finding unequivocally establish that the correct structure of oxybenzotriazole based coupling reagents both in the solution and solid states are benzotriazolyl N-oxides as shown in la, 2a and 3a and not the uronium form as represented in the current literature
[Back to top] SIMPLE AND EFFICIENT METHOD FOR SYNTHESIS OF Z-/Boc-AMINO ACID AMIDES USING p-TOLUENESULPHONYL CHLORIDE
Kuppanna
Ananda, Ganga-Ramu Vasanthakumar and Vommina V. Suresh Babu
Amides of Z- and Boc protected
amino acids have been prepared by using p-to1uenesu1phonyl chloride (TsCl) for the activation of carboxyl group. The
resulting mixed carboxylic-sulphonic anhydride intermediate was treated in situ with an excess of 25% ammonia
solution. All the amino acid amides prepared were obtained as crystalline solids in good yield and purity.
[Back to top] CD CONFORMATIONAL AND MODELING STUDIES OF A SYNTHETIC PEPTIDE VPDLLADLLK IN DIFFERENT MEDIA
J.Shobin, A.K.Mishra, & Nagasuma Chandra
CD spectral studies of
VPDLLADLLK, a synthetic peptide shows that it undergoes a conformational
transition from an unordered structure to
a more ordered structure from a polar to a non-polar homogeneous medium. In microheterogeneous media like SDS, CT AB
micelles and DMPC lipid bilayer, the peptide exhibits a more stable a- helical structure. The helical
conformation is stabilized in DMPC lipid bilayer. Homology modeling gives the picture of a- helix, where the
middle six residues LLADLL form the turns of the helix.
[Back to top] CRYSTALLIZATION AND PRELIMINARY X-RAY STUDIES OF L-(+)-2,3-BUTANEDIOL DEHYDROGENASE FROM BREVIBACTERIUM SACCHAROLYTICUMC-I012
Masato
Otagiri, Genji Kurisu, Sadaharu Ui , Moriya Ohkuma, Toshiaki Kudo and Masami
Kusunoki
L(+)-2,3-Butanediol dehydrogenase
(L-BDH) from Brevibacterium saccharolyticum C-1012 has been crystallized by the hanging drop vapor
diffusion method with polyethylene glycol 4000 as the precipitant. Crystals of L-BDH belong to the triclirlic
system, space group PI with cell dimensions a = 60.8 Å, b = 69.2 Å, c = 127.4 Å, a = 96.1°, β= 100.2° and γ
= 109.6°. Crystals diffracted to 2.0 Å resolution on a synchrotron radiation, and a full data set was collected
at resolution of 2.0 Å at 100K.
[Back to top] ULTRA-HIGH RESOLUTION X-RA Y DIFFRACTION FROM CRYSTALS OF THE KINETIC MUTANT OF HUMAN CARBONIC ANHYDRASE II, HIS 64 ALA, AND ITS COMPLEXES WITH PROTON ACCEPTOR/DONORS
David
Duda, Chingkuang Tu, David N. Silverman, A. JosephUJHh Kalb (Gilboa) Mavis
Agbandje-McKenna:, Robert McKenna
Crystals of human carbonic anhydrase II with a specific point mutation, His 64 to Ala, have been grown in a soiution of ammonium sulfate in the presence of mercury chloride. The crystals appear in approximately two weeks and belong to the monoclinic space group P21, with unit cell parameters of a = 42.2 Å, b = 41.4 Å, c = 71.9 Å, β = 1042° and one carbonic anhydrase molecule in the asymmetric unit. The crystals diffract X rays beyond 1.0 Å resolution. These crystals, soaked with exogenous proton acceptor/donors, will be used in X-ray and neutron diffraction studies to map the fine water structure "proton wire" in the active site of carbonic anhydrase and to assign the intra- and intermolecular proton transfer pathway(s) from the zinc bound water out to the bulk solvent
[Back to top] CRYSTALLIZATION AND PRELIMINARY X-RAY DIFFRACTION STUDY OF RIBOFLA VIN SYNTHASE
Zdzislaw
Wawrzak, Paul V. Viitanen, Joseph C. Calabrese, and Douglas B. Jordan
Escherichia coli riboflavin
synthase "crystallizes at 22 °C in the presence of 7-10% by volume
diglyme, 20-50 mM MgCl2 and pH 7.0. In this medium diffraction quality crystals
are routinely obtained within 5 h and they are stable for 10 weeks. The
crystals are orthogonal in space group P2l2l21
with unit cell dimensions of a = 52.4Å, b = 62.1 Å, c = 218.8 Å. A 97% complete
data set was collected at 2.1 Å resolution.