Water in Crystal Contacts: Retention of Bridging Waters. Pp. 80-86.
Travis Gallagher and Gary L.Gilliland
[Abstract]
An O-Phosphotyrosine-Containing Analogue of Human Angiotensin II Exhibits
Increased Serum Stability. Pp. 87-90.
John D. Wade, John W Perich, Geoffrey W. Tregear, Gyorgyi I. Szendrei and
Laszlo Otvos, Jr.
[Abstract]
Calculation of the Proton Chemical Shifts as a Tool to Explain the Peculiarities of 1H NMR Spectra of Protein in a Compact Denatured State. Pp. 91-98.
Lilly S. Kivaeva, Gulshat T. Ibragimova and Victor P. Kutyshenko
[Abstract]
Characterization of the Minor Impurities During Synthesis of Alzheimer beta-Protein. Pp. 99-106.
Sandip B. Vyas and Lawrence K. Duffy
[Abstract]
Solution Structure of the Wild-Type HMG1 A Box by 1H and 15N NMR and
Molecular Modelling. Pp. 107-114.
Y. Ohyama, P. Sodano, D. Locker, M.E. Bianchi, M. Leng, F. Vovelle and M. Ptak
[Abstract]
Intestinal Absorption of Lactonolactone-Coupled Tripeptide and Stability in Serum: Effect of Linkage Between Peptide and Monosaccharide Moiety. Pp. 115-122.
Takashi Mizuma, Kunihiro Ohta and Shoji Awazu
[Abstract]
Temperature Effect on the Structural Features of beta-Glycosidase from Sulfolobus solfataricus. An Infrared Study. Pp. 123-130.
Sabato D'Auria, Mose Rossi, Roberto Nucci, Enrico Bertoli and Fabio Tanfani
[Abstract]
The Primary Strucutre of the beta Subunit of Desulfovibrio desulfuricans (ATCC 27774) [NiFe] Hydrogenase. Pp. 131-138.
Ricardo Franco, Juan J. Calvete, Hubert H. Thole, Manfred Raida, Isabel Moura and José J.G. Moura
[Abstract]
Crystallisation Reports
Crystallization and Preliminary X-Ray Analysis of Triosephosphate Isomerase from
Trypanosoma cruzi. Pp. 139-144.
Ernesto Maldonado, Abel Moreno, Kaliyamoorthy Penneerselvarn, Pedro Ostoa-Saloma, Georgina Garza-Ramos, Manuel Soriano-García, Ruy Pérez-Montfort,
Marietta Tuena de Gómez-Puyou and Armando Gómez-Puyou
[Abstract]
Crystallization and Preliminary X-Ray Crystallographic Studies of dUTPase from
Equine Infectious Anemia Virus. Pp. 145-148.
Rebecca Persson, Anna Maria Rosengren, Per Olof Nyman, Zhigniew Dauter and
Eila Cedergren-Zeppezauer
[Abstract]
[Back to top] E. coli Expression of Functionally Active Cellular Targeting Motif of Human Focal Adhesion Kinase. Ryuji Yamaguchi, Hisato Jingami and Judith M. Healy.
Focal adhesion kinase (FAK) is a structurally distinctive cytoplasrnic tyrosine kinase which 1ocalizes to cellular focal contacts. Sequences near the C-terminus of FAK, termed the focal adhesion targeting (FAT) domain, are responsible for localization of FAK to focal adhesion complexes. Here, we report expression in E. coli of C-terminal fragments of human FAK containing the FAT domain as epitope-tagged fusions. Purified bacterially derived FAT domains were biologically active and their far ultraviolet circular dichroism spectra showed evidence of extensive alpha-helical secondary structure.
[Back to top] Water in Crystal Contacts: Retention of Bridging Waters. Travis Gallagher and Gary L. Gilliland.
In 3 well-refined crystal structures of subtilisin BPN' in different space groups, crystal contact hydration was compared. The structures are: 1sup, space group C2, resolution 1.6 A, with 194 waters; 2st 1 , P2/1 2/1 2/1, resolution 1.8 A, with 154 waters, and 1sub, P2/1, resolution 1.75 A, with 200 waters. Of 106 bridging water sites, approximately 50% retain the water in the absence of the crystal contact, suggesting that bound water may serve as a template for crystal contact formation. Bridging waters that are retained on both surfaces must overlap when the contact forms, eliminating one water in a condensation reaction. In such cases, the water sites are geometric determinants of the contact. Specific examples are described in the C2 crystal structure.
[Back to top] An O-Phosphotyrosine-Containing Analogue of Human Angiotensin II Exhibits
Increased Serum Stability. John D. Wade, John W Perich, Geoffrey W. Tregear, Gyorgyi I. Szendrei and Laszlo Otvos, Jr.
Synthetic human [Tyr (P)]4-angiotensin II was analyzed by CD spectroscopy and compared with native human angiotensin II. The data showed there to be no apparent conformational difference between the two peptides in both aqueous and graded TFE solutions. All assessment of the in vitro stability of the two peptides to serum proteases showed clearly that the 0-phosphorylated angiotensin II was more resistant to enzymic degradation suggesting that the tyrosyl 0-phosphate plays a significant role in inhibiting this process.
[Back to top] Calculation of the Proton Chemical Shifts as a Tool to Explain the Peculiarities of 1H NMR Spectra of Protein in a Compact Denatured State. Lilly S. Kivaeva, Gulshat T. Ibragimova and Victor P. Kutyshenko.
Using the protein crystal structure the chemical shift dispersions of binase alpha-CH protons were calculated for protein in native state and in a compact denatured one. The same models of internal fields with different adjusting parameters were used to describe data for protons disposed in regions of both regular and irregular secondary structures. It was shown that peculiarities of H NMR spectra observed for compact denatured proteins could be explained through fluctuations of hydrogen bonds network in conserved secondary structures.
[Back to top] Characterization of the Minor Impurities During Synthesis of Alzheimer beta-Protein. Sandip B. Vyas and Lawrence K. Duffy.
Synthetic Alzheimer beta-protein, beta1-40, was analyzed for minor impurities after assembly from activated esters of fmoc-amino acids. Analytical RPHPLC show that beta26-40 and beta8-40 were the most common termination peptides. Co-elution of hetero-oligomeric protein species resulted in band broadening on reversed-phase chromatography. Data on co-incubation of pure peptides indicate that a kinetic barrier had to be overcome for hetero-oligomerization. The presence of these peptides may affect the reproducibility of aggregation and toxicity assays.
[Back to top] Solution Structure of the Wild-Type HMG1 A Box by 1H and 15N NMR and Molecular Modelling. Y. Ohyama, P. Sodano, D. Locker, M.E. Bianchi, M. Leng, F. Vovelle and M. Ptak.
The solution structure of the wild-type A box of rat HMG1 has been determined by heteronuclear three-dimensional NMR and molecular modelling. This structure adopts an "L" shape structure globaly similar to that described for a mutant A box and other HMO boxes. However the wild-type A box contains two Cys residues which could form an intramolecular bridge in an oxidative environment. This should be into competition with a contribution of the thiol groups in the binding of HMG proteins to cisplatin-modified DNA.
[Back to top] Intestinal Absorption of Lactonolactone-Coupled Tripeptide and Stability in Serum: Effect of Linkage Between Peptide and Monosaccharide Moiety. Takashi Mizuma, Kunihiro Ohta and Shoji Awazu.
Aminopeptidase-degradable tyrosylglycylglycine (TGG), a part of leucine enkephalin, was coupled with lactonolactone to conjugate galactose with TGG by way of amide bond linkage. Lactonolactone-coupling improved the intestinal absorption of TGG as well as the stability in intestine and in serum, like galactose conjugation by way of CH2-NH linkage (lactose-coupling). However, lactonolactone-coupling also provided TGG with peptidase-susceptible property different from that of TGG.
[Back to top] Temperature Effect on the Structural Features of beta-Glycosidase from Sulfolobus solfataricus. An Infrared Study. Sabato D'Auria, Mose Rossi, Roberto Nucci, Enrico Bertoli and Fabio Tanfani.
In this paper we report the temperature effect on the structural properties of a thermostable beta-glycosidase isolated from archaeon Sulfolobus solfatancus monitored by infrared spectroscopy. The investigations were carried out at pH 10, because at neutral pH's values the denaturation process did not take place. The denaturation process was irreversible and sodium dodecyl sulfate additions to the enzyme solution caused a lower temperature and a lower onset of the protein denaturation.
[Back to top] The Primary Strucutre of the beta Subunit of Desulfovibrio desulfuricans (ATCC 27774) [NiFe] Hydrogenase. Ricardo Franco, Juan J. Calvete, Hubert H. Thole, Manfred Raida, Isabel Moura and José J.G. Moura.
The periplasmic [NiFe] hydrogenase isolated from Desulfovibrio (D.) desulfuricans (ATCC 27774) is a heterodimer of a 28 kDa (beta) and a 60 kDa (alpha) subunit. Here we report the complete amino acid sequence of the small (beta) polypeptide chain determined by Edman degradation of proteolytic fragments. Electrospray-ionization mass spectrometry of the native protein confirmed the sequencing results. The sequence is compared with that of D. gigas [NiFe] hydrogenase whose three-dimensional structure has been recently published.
[Back to top] Crystallization and Preliminary X-Ray Analysis of Triosephosphate Isomerase from Trypanosoma cruzi. Ernesto Maldonado, Abel Moreno, Kaliyamoorthy Penneerselvarn, Pedro Ostoa-Saloma, Georgina Garza-Ramos, Manuel Soriano-García, Ruy Pérez-Montfort,
Marietta Tuena de Gómez-Puyou and Armando Gómez-Puyou.
Recombinant triosephosphate isomerase from Trypanosoma cruzi was crystallized at room temperature from 0.1 M Na Hepes pH 7.5, 2 % (v/v) PEG 400 and 2.0 M ammonium sulfate. Crystals belong to the orthorhombic space group P2/1 2/1 2/1 with cell dimensions a = 43.7l A, b = 77.65A, and c = 149.54A, and alpha = beta = gamma = 90 degrees. Another crystal with cell dimensions a = b = 56. 16A, c = 161.85A, and alpha = beta = gamma = 90 degrees with two possible tetragonal space groups P4/1 2/1 2 or P4/3 2/1 2 was obtained. Both crystals diffract up to 1.86 A.
[Back to top] Crystallization and Preliminary X-Ray Crystallographic Studies of dUTPase from Equine Infectious Anemia Virus. Rebecca Persson, Anna Maria Rosengren, Per Olof Nyman, Zhigniew Dauter and Eila Cedergren-Zeppezauer.
Deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase), an enzyme in the nucleotide metabolism, froin the retrovirus Equine Infectious Anemia Virus (EIAV) has been crystallized from PEG mixtures at neutral pH using a vapour diffusion method. Crystals of native enzyme belong to the space group R32, while crystals formed in the presence of Sr2+ and dUDP, a substrate analogue, belong to the space group P4/1 32 or P4/3 32. Both forms are suitable for high resolution X-ray structure analyses.