1984
DOI: 10.1021/bi00297a019
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Resonance Raman carbonyl frequencies and ultraviolet absorption maxima as indicators of the active site environment in native and unfolded chromophoric acyl-.alpha.-chymotrypsin

Abstract: The imidazole of chromophoric p-(dimethylamino)benzoic acid, DABIm, reacts with the serine protease alpha-chymotrypsin in the pH range of 4-7 to form a stable acyl intermediate that gives very good resonance-enhanced Raman spectra. The resonance Raman and absorption spectra of the acyl enzyme intermediate have been compared with the spectra of simple model compounds such as the corresponding chromophoric methyl ester, aldehyde, and imidazole. The resonant Raman and ultraviolet absorption spectra of these simpl… Show more

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Cited by 16 publications
(13 citation statements)
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References 15 publications
(27 reference statements)
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“…Recently Argade et al (1984a) have studied the Raman spectrum of p-dimethylaminobenzoyl-chymotrypsin using 457 nm excitation. In Fig.…”
Section: Resonance Raman Spectroscopymentioning
confidence: 99%
See 1 more Smart Citation
“…Recently Argade et al (1984a) have studied the Raman spectrum of p-dimethylaminobenzoyl-chymotrypsin using 457 nm excitation. In Fig.…”
Section: Resonance Raman Spectroscopymentioning
confidence: 99%
“…, chymotypsin(b) and the difference between the two (c) Spectra were recorded using 458 nm laser excitation, the enzyme being 2.8 mm in each case. Adapted fromArgade et al (1984a).…”
mentioning
confidence: 99%
“…The dimethylaminobenzoyl group has been used before with success to obtain resonance enhanced Raman spectra of enzyme-substrate intermediates in enzyme systems other than CPA (19). This compound is hydrolyzed by CPA to give ß-phenyllactic acid and pdimethylaminobenzoic acid.…”
Section: Summary Of Results On the Time Resolved Far Ultraviolet Ramamentioning
confidence: 99%
“…In addition, Raman spectra of the substrate DABPLA and of its chromophoric product of hydrolysis, DAB acid, indicate that the only band which might occur in this region is the carbonyl band from unionized DAB acid at 1715 Raman spectra of enzyme-substrate intermediates in enzyme systems other than CPA (19) . This compound is hydrolyzed by CPA to give ^-phenyllactic acid and pdimethylaminobenzoic acid.…”
Section: Summary Of Results On the Time Resolved Far Ultraviolet Ramamentioning
confidence: 99%
“…Several physical observations of acyl-enzyme intermediates features of these activated esters. These include an X-ray crystallographic determination of the structure of indoleacryloyl-a-chymotrypsin at 2.5 Á (Henderson, 1970), absorption spectrophotometry investigations of arylacryloyl-achymotrypsins (Bender et al, 1962;Bernhard et al, 1965; Charney & Bernhard, 1967; Bernhard & Malhotra, 1974), resonance Raman studies of acyl-a-chymotrypsins (MacClement et al, 1981;Phelps et al, 1981;Argade et al, 1984), carbon-13 NMR observations of acetyl-a-chymotrypsin (Nie et al, 1977;Tobias et al, 1984) and 5-n-propyl-2-furoyl-achymotrypsin (McWhirter et al, 1985), and fluorine-19 NMR observation of />-(trifluoromethyl)cinnamoyl-and o-, m-, and p-fluorocinnamoyl-a-chymotrypsins (Maddox et al, 1975;Gerig & Halley, 1981). Herein we report that selenium-77 NMR spectroscopy is particularly useful in the characterization of an appropriately substituted selenium-containing acyl-enzyme intermediate.…”
mentioning
confidence: 99%