title: Purification and properties of nuclease SP.
abstract: Single-strand-specific nucleases are a diverse and important group of enzymes that are able to cleave a variety of DNA structures present in duplex molecules. Nuclease SP, an enzyme from spinach, has been purified to apparent homogeneity, allowing for the unambiguous characterization of a number of its physical properties as well as its DNA strand cleavage specificities. The effects of ionic strength, pH, divalent metal cations, and temperature on nuclease SP activity have been examined in detail. Nuclease SP was found to be quite thermostable and could be stimulated by Co2+. In addition, the cleavage of UV-damaged and undamaged supercoiled plasmid substrates under a variety of conditions suggests that at least two types of structures are recognized and processed by nuclease SP: UV photoproduct-induced distortions and unwound "nuclease hypersensitive sites". These studies indicate that nuclease SP is functionally related to other single-strand-specific nucleases and is a potential enzymatic tool for probing and manipulating various types of DNA structures.
authors.affiliation: Department of Biochemistry, School of Medicine, Emory University, Atlanta, Georgia 30322.
authors.forename: J A
authors.initials: JA
authors.lastname: Strickland
authors.forename: L G
authors.initials: LG
authors.lastname: Marzilli
authors.forename: J M
authors.initials: JM
authors.lastname: Puckett
authors.forename: P W
authors.initials: PW
authors.lastname: Doetsch
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000737:chemistry / Q000302:isolation & purification*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008670:Metals / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012492:Salts / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D009711:Nucleotides; D010940:Plant Proteins; D012492:Salts; D004720:Endonucleases; C071228:nuclease SP