RESUMO
The acute massive hemorrhage (35-37% of the blood volume) at rats is accompanied by changes of morphological (diameter, the area, polarizations, the form-factor, integrated and specific absorbency) and biophysical (a relief of a surface and microviscosity of a lipid phase of plasmolemma) characteristics of erythrocytes. Thus character and dynamics of response of erythron initially intact and vagotomized (14 days after operation) animals essentially differ: the former demonstrate significant changes in 3-10 h and 240 h and the latter--in 0.5 h and 96 h.
Assuntos
Eritrócitos/patologia , Hemorragia/patologia , Vagotomia/efeitos adversos , Animais , Membrana Celular/patologia , Hemorragia/etiologia , Masculino , RatosRESUMO
We overexpressed duplex-specific nuclease (DSN) from Kamchatka crab in Escherichia coli cells and developed procedures for purification, renaturation, and activation of this protein. We demonstrated identity of the properties of the native and recombinant DSN. We also successfully applied the recombinant DSN for full-length cDNA library normalization.
Assuntos
Braquiúros/enzimologia , Endonucleases/metabolismo , Proteínas Recombinantes/metabolismo , Animais , Sequência de Bases , Braquiúros/genética , Braquiúros/metabolismo , Clonagem Molecular/métodos , DNA Complementar/genética , Endonucleases/química , Endonucleases/genética , Ativação Enzimática , Estabilidade Enzimática , Ácidos Nucleicos Heteroduplexes/metabolismo , Dobramento de Proteína , Renaturação Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
Antimicrobial peptides (AMPs), named lycocitin 1, 2 and 3, and a peptide with a monoisotopic molecular mass of 3038.70 Da were detected in the venom glands of the wolf spider Lycosa singoriensis. Two of the peptides, lycocitin 1 and 2, are new AMPs whereas lycocitin 3 is highly homologous to lycotoxin II isolated from the venom of spider Lycosa carolinensis. In addition, two other peptides with monoisotopic masses of 2034.20 and 2340.28 Da showing the motif typical for antimicrobial peptides were also identified. These peptides and lycocitin 1, 2 and 3 were de novo sequenced using electron capture dissociation and low-energy collisional tandem mass spectrometry. The amino acid sequence of lycocitin 1 was determined as GKLQAFLAKMKEIAAQTL-NH(2). Lycocitin 2 differs from lycocitin 1 by a replacement of a lysine residue for an arginine residue at the second position. Lycocitin 3 differs from the known lycotoxin II consisting of 27 amino acid residues by a deletion of Gly-26. Both lycocitin 1 and 2 inhibit growth of Gram-positive (Staphylococcus aureus, Bacillus subtilis) and Gram-negative (Escherichia coli) bacteria and fungi (Candida albicans, Pseudomonas aeruginosa) at micromolar concentrations.
