Details

Autor(en) / Beteiligte

• Funakoshi, A
• Tsubota, Y
• Fujii, K
• Ibayashi, H
• Takagi, Y

Titel

Simple purification and properties of bovine pancreatic deoxyribonuclease I

Ist Teil von

• Journal of biochemistry (Tokyo), 1980-10, Vol.88 (4), p.1113-1118

Ort / Verlag

England: Oxford University Press

Erscheinungsjahr

1980

Quelle

MEDLINE

Beschreibungen/Notizen

• A simple purification method for pancreatic deoxyribonuclease I (DNase I) EEC 3.1.4.3] was developed by utilizing the technique of isoelectric focusing. The active protein was resolved into at least four forms with different isoelectric points; the major components a, b, and c had isoelectric points at pH 5.2, 4.9, and 4.8, respectively, and that of the minor component d was at 4.7. The four components (a, b, c, and d) exhibited peaks similar to those observed by Salnikow el al. after phosphocellulose chromatography (A, B, C, and D). The four components were all free from RNase and protease activities and were very stable at 0–2°C for at least four weeks. Further, each of the four peaks exhibited a single protein band after polyacrylamide electrophoresis. DNase I-a antibody was prepared; it was very specific for DNase I and precipitated with the other components (b, c, and d). The mode of endonucleolytic action of pancreatic DNase I-a purified from Worthington DP grade DNase I was investigated. The sedimentation patterns in neutral sucrose gradients of digest of circular duplex DNA in an early stage of hydrolysis suggested that DNase I produces single strand scissions in the initial attack in the presence of divalent metal ions.