Дезоксирибонуклеаза I

Использование

Deoxyribonuclease I is used for the removal of DNA from protein samples. DNAse I is used to nick DNA as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used during the isolation of plasma membrane vesicles from Neurospora crassa cell culture.1 It has also been used along with trypsin for the preparation of single cell suspension from rat testes. Deoxyribonuclease I from bovine pancreas has been used in a study to compare several procedures for reducing RNase contamination in preparations of DNase. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effect of the composition of sodium dodecyl sulfate preparations on the renaturation of enzymes after polyacrylamide gel electrophoresis

Общее описание

DNase I, or Deoxyribonuclease I, is an endonuclease isolated from bovine pancreas.

• Our DNase I Digests double str and single stranded DNA into oligo and mononucleotides.
• Bovine pancreatic DNase exists as four isozymes, having isoelectric points for A, B, C and D: 5.22, 4.96, 5.06 and 4.78.3. The predominant form is A, with smaller amounts of B and C, and only minor amount of D.
• DNase I structure resembles the structure of to exonuclease III. It includes two central ? sheets. Each β sheet is composed of six β-strands. This complex of β sheets is surrounded by extensive loop and α-helical regions. This enzyme shares structural similarity to exonuclease III.^[1]

Клиническое использование

DNAse is a human endonuclease, normally present insaliva, urine, pancreatic secretions, and blood. The enzymecatalyzes the hydrolysis of extracellular DNA into oligonucleotides.Aerosolized recombinant human deoxyribonucleaseI (rhDNAse), dornase alfa, Pulmozyme, has beenformulated into an inhalation agent for the treatment of pulmonarydisease in patients with CF.Among the clinical manifestations of CF are obstructionof the airways by viscous, dehydrated mucus. Pulmonaryfunction is diminished, and microbes can become entrappedin the viscid matrix. A cycle of pulmonary obstruction andinfection leads to progressive lung destruction and eventualdeath before the age of 30 for most CF patients. The immunesystem responds by sending in neutrophils, and theseaccumulate and eventually degenerate, releasing largeamounts of DNA. The high levels of extracellular DNA releasedand the mucous glycoproteins are responsible for thedegenerating lung function. The DNA-rich secretions alsobind to aminoglycoside antibiotics typically used to treat theinfections. In vitro studies showed that the viscosity of thesecretions could be reduced by application of DNAse I.
Pulmozyme is approved for use in the treatment of CFpatients, in conjunction with standard therapies, to reducethe frequency of respiratory infections requiring parenteralantibiotics and to improve pulmonary function. The dose isdelivered at a level of 2.5 mg daily with a nebulizer.Pulmozyme is not a replacement for antibiotics, bronchodilators,and daily physical therapy.