ABSTRACT
Metacaspases, cysteine proteases belonging to the peptidase C14 family, are suspected of being involved in the programmed cell death of plants, although their sequences and substrate specificity differ from those of animal caspases. At present, the knowledge on the metacaspase reaction mechanism is based only on biochemical data and homology models constructed on caspase templates. Here we propose a novel template for metacaspase modeling and demonstrate important advantages in comparison to the conventionally used caspase templates. We also point out the connection between plant and bacterial metacaspases, underlining the prokaryotic roots of Programmed Cell Death (PCD).
Subject(s)
Caspases/chemistry , Geobacter/enzymology , Sequence Analysis, Protein/methods , Sequence Homology, Amino Acid , Triticum/enzymology , Algorithms , Amino Acid Sequence , Bacterial Proteins/chemistry , Caspase 7/chemistry , Catalytic Domain , Cell Death , Databases, Protein , Humans , Models, Molecular , Molecular Sequence Data , Plant Proteins/chemistry , Protein Folding , Protein Structure, Secondary , Sequence Alignment , Substrate SpecificityABSTRACT
Cloning using bacterial artificial chromosomes (BACs) can yield high quality genomic libraries, which are used for the physical mapping, identification and isolation of genes, and for gene sequencing. A BAC genomic library was constructed from high molecular weight DNA (HMW DNA) obtained from nuclei of the cucumber (Cucumis sativus L. cv. Borszczagowski; B10 line). The DNA was digested with the HindIII restriction enzyme and ligated into the pCC1BAC vector. The library consists of 34,560 BAC clones with an average insert size of 135 kb, and 12.7x genome coverage. Screening the library for chloroplast and mitochondrial DNA content indicated an exceptionally low 0.26% contamination with chloroplast DNA and 0.3% with mitochondrial DNA.
Subject(s)
Chromosomes, Artificial, Bacterial/genetics , Cucumis sativus/genetics , Gene Library , Chromosome Mapping , Cloning, Molecular/methods , DNA, Plant/genetics , DNA, Plant/isolation & purification , DNA, Plant/metabolism , Electrophoresis, Gel, Pulsed-Field , Escherichia coli/genetics , Genetic Vectors , Molecular Weight , Plant Leaves/genetics , Transformation, BacterialABSTRACT
These studies were focused on the influence of two treatment methods of patients with chronic renal disease on RBC deformability. In peritoneal dialysed-patients (PD) erythrocytes exhibited a decrease in their deformability as compared to control subjects whereas this parameter in RBC deriving from hemodialysed patients (HD) was not altered. The alleviating effect of plasma components on deformability of erythrocytes was confirmed after the isolation of a pure faction of these cells as the parameter became worse. The activity of studied enzymes: acetylcholinesterase (Ache), dehydrogenase glucose-6-phosphate (G-6-PD) and glutathione reductase (GR) maintained their physiological values in both dialysis groups.
