1973
DOI: 10.1021/bi00748a006
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Interaction of guanine ligands with ribonuclease T1

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Cited by 35 publications
(44 citation statements)
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“…The dissociation constant for 3'-GMP and barnase was measured using UV difference spectroscopy. The UV difference spectrum of the barnase-3'-GMP complex relative to free barnase and 3'-GMP has a maximum at 290 nm and a minimum at 243 nm, similar to that observed previously for RNase T1 (Walz & Hooverman, 1973). The difference in absorbance at 290 and 243 nm ( = 290 -AAj4z) was measured upon titrating a solution of barnase (20>iM protein, 59.8 mM acetic acid/40.2 mM sodium acetate, pH 4.5) with a concentrated solution of 3'-GMP [disodium salt (Sigma) in H2O].…”
Section: Methodssupporting
confidence: 86%
“…The dissociation constant for 3'-GMP and barnase was measured using UV difference spectroscopy. The UV difference spectrum of the barnase-3'-GMP complex relative to free barnase and 3'-GMP has a maximum at 290 nm and a minimum at 243 nm, similar to that observed previously for RNase T1 (Walz & Hooverman, 1973). The difference in absorbance at 290 and 243 nm ( = 290 -AAj4z) was measured upon titrating a solution of barnase (20>iM protein, 59.8 mM acetic acid/40.2 mM sodium acetate, pH 4.5) with a concentrated solution of 3'-GMP [disodium salt (Sigma) in H2O].…”
Section: Methodssupporting
confidence: 86%
“…The RNase Ti was a gift of Professor F. G. Walz of Kent State University, prepared by the method of Hooverman (27). Gel electrophoresis exhibited a single band, and the UV absorption spectrum showed that there was no observable contamination.…”
Section: Methodsmentioning
confidence: 99%
“…GpcU binding with RNase T 1 . Ribonuclease T 1 (Gln25 allozyme) was purified as previously described and its concentration was determined with a molar absorptivity at 278 nm ( 278 nm ) of 2.1 × 10 4 M -1 cm -1 (24). The concentration of GpcU was determined with the molar absorptivity for GpU [ 280 nm ) 1.06 × 10 4 M -1 cm -1 (25)].…”
Section: Preparation Of Gpcumentioning
confidence: 99%
“…The concentration of GpcU was determined with the molar absorptivity for GpU [ 280 nm ) 1.06 × 10 4 M -1 cm -1 (25)]. Binding was studied at pH 5.5 using ultraviolet difference spectroscopy (24). The maximum difference absorbance occurred at 291 nm and values of the difference absorbance, ∆A, at this wavelength were determined for a fixed total concentration of the enzyme ([E] 0 ) 3.43 × 10 -3 M) in the presence of variable total concentrations of GpcU ([L] 0 ; (0.479-2.21) × 10 -4 M).…”
Section: Preparation Of Gpcumentioning
confidence: 99%