Active site histidine in spinach ribulosebisphosphate carboxylase/oxygenase modified by diethyl pyrocarbonate

Biochemistry ◽  
1985 ◽  
Vol 24 (15) ◽  
pp. 3957-3962 ◽  
Author(s):  
Yasuo Igarashi ◽  
Bruce A. McFadden ◽  
Ted El-Gul
Keyword(s):  
Active Site ◽  
Diethyl Pyrocarbonate ◽  
Ribulosebisphosphate Carboxylase ◽  
Active Site Histidine

scholarly journals Single active-site histidine in d-xylose isomerase from Streptomyces violaceoruber. Identification by chemical derivatization and peptide mapping

1989 ◽  
Vol 263 (1) ◽  
pp. 195-199 ◽  
Author(s):  
W Vangrysperre ◽  
C Ampe ◽  
H Kersters-Hilderson ◽  
P Tempst
Keyword(s):  
Active Site ◽  
Peptide Mapping ◽  
Xylose Isomerase ◽  
Specific Chemical ◽  
Peptide Cleavage ◽  
Diethyl Pyrocarbonate ◽  
Streptomyces Violaceoruber ◽  
Wavelength Detection ◽  
Micro Organisms ◽  
Active Site Histidine

Group-specific chemical modifications of D-xylose isomerase from Streptomyces violaceruber indicated that complete loss of activity is fully correlated with the acylation of a single histidine. Active-site protection, by the ligand combination of xylitol plus Mg2+, completely blocked diethyl pyrocarbonate derivatization of this particular residue [Vangrysperre, Callens, Kersters-Hilderson & De Bruyne (1988) Biochem. J. 250, 153-160]. Differential peptide mapping between D-xylose isomerase, which has previously been treated with diethyl pyrocarbonate in the presence or absence of xylitol plus Mg2+, allowed specific isolation and sequencing of a peptide containing this active-site histidine. For this purpose we used two essentially new techniques: first, a highly reproducible peptide cleavage protocol for protease-resistant, carbethoxylated proteins with guanidinium hydrochloride as denaturing agent and subtilisin for proteolysis; and second, reverse-phase liquid chromatography with dual-wavelength detection at 214 and 238 nm, and calculation of absorbance ratios. It allowed us to locate the single active-site histidine at position 54 in the primary structure of Streptomyces violaceoruber D-xylose isomerase. The sequence around this residue is conserved in D-xylose isomerases from a diversity of micro-organisms, suggesting that this is a structurally and/or functionally essential part of the molecule.

Identification of an active site histidine in urokinase

1976 ◽  
Vol 429 (1) ◽  
pp. 252-257 ◽  
Author(s):  
Eng Bee Ong ◽  
Alan J. Johnson ◽  
Guenther Schoellmann
Keyword(s):  
Active Site ◽  
Active Site Histidine

scholarly journals The location of the active-site histidine residue in the primary sequence of papain

1968 ◽  
Vol 108 (5) ◽  
pp. 861-866 ◽  
Author(s):  
S. S. Husain ◽  
G. Lowe
Keyword(s):  
Amino Acid ◽  
Sodium Borohydride ◽  
Active Site ◽  
Iodoacetic Acid ◽  
Cysteine Residue ◽  
Histidine Residue ◽  
Primary Sequence ◽  
Active Site Cysteine ◽  
Active Site Histidine ◽  
Active Site Cysteine Residue

Papain that had been irreversibly inhibited with 1,3-dibromo[2−14C]acetone was reduced with sodium borohydride and carboxymethylated with iodoacetic acid. After digestion with trypsin and α-chymotrypsin the radioactive peptides were purified chromatographically. Their amino acid composition indicated that cysteine-25 and histidine-106 were cross-linked. Since cysteine-25 is known to be the active-site cysteine residue, histidine-106 must be the active-site histidine residue.

Sign in / Sign up

Export Citation Format

Share Document