Protein and Peptide Letters, Vol. 7, No. 4, 2000
Conformation of the a-Aminoacrylate Intermediate of O-Acetylserine sulfhydrylase from Salmonella typhimurium as Inferred by 31P NMR Spectroscopy. Pp. 207-210.
Chia-Hui-Tai, Paul F. Cook and Klaus D. Schnackerz*
Simulation of the Highly Stable Protein: Bovine gB-Crystallin at Room and High Temperature. Pp. 211-217.
Andrew G.
Purkiss, Christine Slingsby and Julia M. Goodfellow*
Sensitive Fluorogenic Substrates for Plasmepsin 21. Pp.219-223.
Sergei V.
Gulnik*, Leonid I. Suvorov, Pavel Majer and John W. Erickson
Effect of a Toxic Protein Isolated from Zea mays seeds on the development and survival of the Copea Weevil, Callosobruchus maculates. Pp. 225-231.
Maria Ligia
Rodrigues Macedo*, Mirela Batista Coelho, Maria das Gracas Machado
Freire, Olga Lima Tavares Machado, Sergio Marangoni and Jose Camillo Novello
A Novel Isoform of Osteoprotegerin Gene: Cloning and Expression and Its Hypocalcemic Effect in Mice*. Pp. 233-240.
Zhi-Yong He,
Guan-Zhen Yang, Zhe-Yu Chen, Biao Li, Wei-Jie Zhang, Xiang-Fu Wu*
Further Characterization of the Glycan-Binding Specificity of the Seed Lectin from Vatairea Macrocarpa and Its Dependence of pH. Pp. 241-248.
Marcio V.
Ramos*, Liezelotte R. Bomfim, Benildo S. Cavada, Nylane M.N.
Alencar, Thalles B. Grangeiro and Henri Debray
Molecular Modeling of Arcelin-5C from Bean Seeds and Determination of Its Solution State. Pp. 249-256.
Artur T.
Cordeiro, Isabel R. Gerhardt, Osmundo B. de Oliveira-Neto, Carlos Bloch Jr. and
Maria Fatima Grossi de sa*
Expression and Purification of Human Metallothionein-3 Beta Domain in E. coli*. Pp. 257-263.
Daqing Wu*,
Jiong Shen, Binggen Ru
Location of Cysteine Residues and the Biological Activities of the Fluorescence-labeled Cinnamoin. Pp. 265-272.
Liang Xie,
Fa-Jian Hou, Wang-yi Liu* and En-duo Wang
Crystallization and Preliminary Crystallographic Studies of Pekin Duck Oxyhemoglobin. Pp. 273-276.
Huan-Chen
Wang, Zheng Zha, Yu-He Liang, Zi-Qian Hua and Guang Ying Lu*
Purification and Crystallization of a Ferredoxin Reductase Component of a Biphenyl Dioxygenase Derived from Pseudomonas sp. strain KKS102. Pp. 277-280.
Takahiro
Yamada, Nobuyuki Sakurai, Tomoko Nishizaki, Toshiya Senda*, Eiji
Masai, Masao Fukuda and Late Yukio
Mitsui
[Back to top] Conformation of the a-Aminoacrylate Intermediate of O-Acetylserine sulfhydrylase from Salmonella typhimurium as Inferred by 31P NMR Spectroscopy.
Chia-Hui-Tai, Paul F. Cook and Klaus D. Schnackerz*
31P NMR chemical shifts of the internal aldimine and the a-aminoacrylate intermediate of O-acetyklserine sulfhydrylase were determined and found to be 5.13 and 3.95 ppm, respectively. In addition, the line width decreases upon addition of OAS, suggesting a less restricted rotation around the C5’-O bond of pyridoxal 5’-phosphate in the a-aminoacrylate intermediate. The upfield shift of the a-aminoacrylate is similar to the behavior of the L-serine external Schiff base and may be interpreted as a loosening of the active site.
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Simulation of the Highly Stable Protein: Bovine gB-Crystallin at Room and High Temperature.
The behavior of the eye-lens protein bovine gB-crystallin is investigated at both room temperature and at an elevated temperature (at which unfolding would be expected to occur) using molecular dynamics. Even in the high temperature simulation, there is little loss of secondary structure. Although only small changes in the tertiary contacts occur, even at high temperature, fluctuations in conformation are much more pronounced at 500K than 300K. these results agree with the experimental observations on the remarkable stability of the eye lens proteins and also indicate where the initial events in unfolding are occurring.
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Sensitive Fluorogenic Substrates for Plasmepsin 21.
Pp.219-223.
Plasmepsin 2 is a malarial aspartic proteinase that has been implicated in initial steps of hemoglobin degradation in parasite and thus represents and attractive antimalarial target. We designed and synthesized a number of quenched fluorogenic substrates and measured the kinetic constants of their cleavage by plasmepsin-2. High sensitivity and superior kinetic properties make the substrates suitable for different application including fluorescence plate reader.
[Back to top] Effect of a Toxic Protein Isolated from Zea mays seeds on the development and survival of the Copea Weevil, Callosobruchus maculates.
Maria Ligia Rodrigues Macedo*,
Mirela Batista Coelho, Maria das Gracas Machado Freire, Olga Lima Tavares Machado,
Sergio Marangoni and Jose Camillo Novello
A new toxic glycoprotein (Zeatoxin) has been isolated from Zea mays and sequenced. The toxin is a single polypeptide chain, with Mr of 10 kDa. Zeatoxin was not digested by a mixture of pepsin and papain or by midgut preparations of C. maculates. A diet of artificial seeds (final concentration, 0.25%) was lethal to 50% of C. maculates. These results shows that Zeatoxin is a new toxic glycoprotein to C. maculates. A search in the database indicated that the N-terminal sequence show no homology to any other known protein.
[Back to top] A Novel Isoform of Osteoprotegerin Gene: Cloning and Expression and Its Hypocalcemic Effect in Mice*.
Zhi-Yong He,
Guan-Zhen Yang, Zhe-Yu Chen, Biao Li, Wei-Jie Zhang, Xiang-Fu Wu*
A novel osteoprotegerin (OPG) isoform cDNA was cloned and expressed in the insect cell and yeast. No homodimeric OPG was found according to the SDS-PAGE analysis and western blot. The OPG isoform was 8 amino acid residues less than the reported hOPG. Genomic DNA of Chinese PBL and HEK 293 cells sequence analysis revealed that the mutation was not the result of RNA editing. The rhOPG with His-Tag at N-terminal did not express the hypocalcemic effect in the normal mice.
[Back to top] Further Characterization of the Glycan-Binding Specificity of the Seed Lectin from Vatairea Macrocarpa and Its Dependence of pH.
Marcio V.
Ramos*, Liezelotte R. Bomfim, Benildo S. Cavada, Nylane M.N.
Alencar, Thalles B. Grangeiro and Henri Debray
The lectin from Vatairea macrocarpa seeds exhibits a pronounced specificity for glycoproteins possessing Gal/GalNAc residues in their carbohydrate moieties. Determination of the fine sugar specificity of the lectin by affinity chromatography of labeled oligosaccharides and glycopeptides of known structures on the immobilized lectin shows that the lectin precisely recognizes non reducing N-actyllactosamine sequence. This interaction seems to depend of the number of N-acetyllactosamine sequence in each structure and is dependent of pH.
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Molecular
Modeling of Arcelin-5C from Bean Seeds and Determination of Its Solution State.
Artur
T. Cordeiro, Isabel R. Gerhardt, Osmundo B. de Oliveira-Neto, Carlos Bloch Jr.
and Maria Fatima Grossi de sa*
Phaseolus arcelin proteins are known to be involved in resistance mechanisms to bruchid pests in plants. The mechanism of action against the larvae is still unclear, but it seems to be related to the oligomeric state and stability of the protein in solution. The solution state of arcelin-5 variants was analyzed by size exclusion chromatography and a molecular model of arcelin-5c was built and compared with the structure of arcelin-1 and phytochemagglutin-L.
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Expression
and Purification of Human Metallothionein-3 Beta Domain in E. coli*.
Daqing Wu*,
Jiong Shen, Binggen Ru
b-domain of metallothionein-3/growth inhibitory factor(MT-3/GIF) is crucial to the neuronal growth inhibitory activity of MT-3. The MT-3 b domain was efficiently expressed as the soluble fusion from with GST protein at a high level which accounted for over 30% total cellular proteins. After the thrombin cleavage and purification, the bioactive recombinant MT-3 b domain was recovered with the yield of 6 mg/L of medium.
[Back to top] Location of Cysteine Residues and the Biological Activities of the Fluorescence-labeled Cinnamoin.
Liang Xie,
Fa-Jian Hou, Wang-yi Liu* and En-duo Wang
Cinnamomin is a type II ribosome-inactivating protein (RIP) isolated from the seeds of Clinnamomum camphora. It is composed of two glycopeptide chains (A- and B-chain). The A-chain exhibits RNA N-glycosidase activity, depurinating an adenosine from the largest ribosomal RNA and thus inhibiting protein synthesis. The B-chain is a lectin, recognizing the galactose-containing receptor on the cell surface and facilitating entry of the A-chain into the cell. A total of ten cysteines were titrated with DTNB in the cinnamomin molecule, one in the A-chain and the other in the B-chain. All the cysteines existed in te disulfide bond form in cinnamomin, which probably accounted for its high structural stability. On the other hand, the sugar chains of cinnamomin were oxidized with periodate and then fluorescence-labeled with FTSC. Both the RNA N-glycosidase activity of it’s a-chain and the lectin activity of its B-chain decreased three fold after fluorescence-labeling.
[Back to top] Crystallization and Preliminary Crystallographic Studies of Pekin Duck Oxyhemoglobin.
Huan-Chen
Wang, Zheng Zha, Yu-He Liang, Zi-Qian Hua and Guang Ying Lu*
Pekin duck oxy hemoglobin has been purified and crystallized using PEG 6000 as precipitant. The crystal is of space group P41212 with lattice constants a = b = 81.23, c = 216.3 Å. There is one tetramer molecule per asymmetric unit. The crystal diffracts up to 2.8 Å resolution and suitable for X-ray crystallographic studies.
[Back to top] Purification and Crystallization of a Ferredoxin Reductase Component of a Biphenyl Dioxygenase Derived from Pseudomonas sp. Strain KKS102.
Takahiro Yamada, Nobuyuki Sakurai, Tomoko Nishizaki, Toshiya Senda*, Eiji Masai, Masao Fukuda and Late Yukio Mitsui
Ferredoxin reducatase component of a biphenyl dioxygenase derived from Pseudomonas sp. Strain KKS102 has been crystallized using sodium formate as the precipitant. The crystal belongs to the space group P6122 (or P6522) with cell parameters of a = b = 99.5 Å, c = 175.6 Å, and diffracts to 2.5 Å resolution.