Preparatin, proteolysis and reversible oxidationof highly purified Azotobacter vinelandii polynucleotide phosphorylase.
1. A new method has been developed for the preparation in good yield of highly purified Azotobacter vinelandii polynucleotide phosphorylase in its reduced form. 2. Aging or digestion with trypsin causes the enzyme to develop a primer requirement that is not eliminated by beta-mercaptoethanol. 3. The development of a primer requirement is accompanied by marked changes of the electrophoretic mobility of the enzyme in polyacrylamide gels. 4. The enzyme is inactivated by aerial oxidation or thiol-specific reagents. The lost activity is restored by beta-mercaptoethanol, but not by oligonucleotide primers.
D000179:Acrylates; D000227:Adenine Nucleotides / Q000378:metabolism; D000388:Air; D001395:Azotobacter / Q000201:enzymology*; D003597:Cytosine Nucleotides / Q000378:metabolism; D004586:Electrophoresis; D005782:Gels; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology; D010406:Penicillins / Q000494:pharmacology; D011108:Polymers; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D013997:Time Factors; D014357:Trypsin
D000179:Acrylates; D000227:Adenine Nucleotides; D003597:Cytosine Nucleotides; D005782:Gels; D010406:Penicillins; D011108:Polymers; D008623:Mercaptoethanol; D012316:RNA Nucleotidyltransferases; D014357:Trypsin