Cholinesterases from flounder muscle. Purification and characterization of glycosyl-phosphatidylinositol-anchored and collagen-tailed forms differing in substrate specificity

1989 ◽  
Vol 181 (3) ◽  
pp. 633-642 ◽  
Author(s):  
Susi STIEGER ◽  
Rene GENTINETTA ◽  
Urs BRODBECK
Keyword(s):  
Substrate Specificity ◽  
Purification And Characterization ◽  
Glycosyl Phosphatidylinositol

scholarly journals Purification and characterization of pig kidney aminopeptidase P. A glycosyl-phosphatidylinositol-anchored ectoenzyme

1990 ◽  
Vol 267 (2) ◽  
pp. 509-515 ◽  
Author(s):  
N M Hooper ◽  
J Hryszko ◽  
A J Turner
Keyword(s):  
Chelating Agents ◽  
Inhibitory Effect ◽  
Purification And Characterization ◽  
Glycosyl Phosphatidylinositol ◽  
Pig Kidney ◽  
Sds Page ◽  
Cyclic Phosphate ◽  
Aminopeptidase P ◽  
Mild Acid

Aminopeptidase P (EC 3.4.11.9) was solubilized from pig kidney membranes with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) and then purified by a combination of anion-exchange and hydrophobic-interaction chromatographies. Contaminating peptidase activities were removed by selective affinity chromatography. The purified enzyme was apparently homogeneous on SDS/PAGE with an Mr of 91,000. Enzymic deglycosylation revealed that aminopeptidase P is a glycoprotein, with up to 25% by weight of the protein being due to the presence of N-linked sugars. The phospholipase-solubilized aminopeptidase P was recognized by an antiserum to the cross-reacting determinant (CRD) characteristic of the glycosyl-phosphatidylinositol anchor. This recognition was abolished by mild acid treatment or deamination with HNO2, indicating that the CRD was due exclusively to the inositol 1,2-cyclic phosphate ring epitope generated by the action of PI-PLC. The activity of aminopeptidase P was inhibited by chelating agents and was stimulated by Mn2+ or Co2+ ions, confirming the metallo-enzyme nature of this peptidase. Selective inhibitors of other aminopeptidases (actinonin, amastatin, bestatin and puromycin) had little or no inhibitory effect.

Purification and Characterization of a Cysteine Endopeptidase fromVasconcellea quercifoliaA. St.-Hil. Latex Displaying High Substrate Specificity

2010 ◽  
Vol 58 (20) ◽  
pp. 11027-11035 ◽  
Author(s):  
M. José Torres ◽  
Sebastián A. Trejo ◽  
M. Inés Martin ◽  
Claudia L. Natalucci ◽  
Francesc X. Avilés ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Purification And Characterization ◽  
High Substrate ◽  
Cysteine Endopeptidase

Purification and characterization of a post-proline dipeptidyl aminopeptidase fromStreptococcus cremorisAM2

1990 ◽  
Vol 57 (1) ◽  
pp. 89-99 ◽  
Author(s):  
Mary Booth ◽  
Ide Ni Fhaoláin ◽  
P. Vincent Jennings ◽  
Gerard O'Cuinn
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Substrate Specificity ◽  
Purification And Characterization ◽  
Ph Optimum ◽  
Cheese Ripening ◽  
Streptococcus Cremoris ◽  
Scissile Bond ◽  
Dipeptidyl Aminopeptidase

SummaryThe present study describes the purification of a post-proline dipeptidyl aminopeptidase from the cytoplasm ofStreptococcus cremorisAM2. On the basis of its elution from a calibrated Sephadex G200 column, the enzyme had a molecular weight of 117000 and exhibited a broad pH optimum activity between 6·0 and 9·0. The activity was most comprehensively inhibited by phenylmethylsulphonylfluoride and more modestly inhibited by 1,10-phenanthroline and 8-hydroxyquinoline but not by EDTA. A range of peptides containing either proline or alanine as the penultimate amino acid residue could act as substrates. The presence of proline on the carboxy side of the scissile bond prevented hydrolysis. However the enzyme could release Pro-Pro from Pro-Pro-Gly-Phe-Ser-Pro. The significance of this substrate specificity is considered in the context of removal of either single proline residues or prolylproline sequences from oligopeptides during cheese ripening.

scholarly journals Purification and characterization of UDP-glucose: hydroxycoumarin 7-O-glucosyltransferase, with broad substrate specificity from tobacco cultured cells

Plant Science ◽  
2000 ◽  
Vol 157 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Goro Taguchi ◽  
Hirofumi Imura ◽  
Yoshio Maeda ◽  
Ritsuko Kodaira ◽  
Nobuaki Hayashida ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Cultured Cells ◽  
Purification And Characterization ◽  
Broad Substrate Specificity

scholarly journals The purification and characterization of acetoacetyl-coenzyme A reductase from Azotobacter beijerinckii

1971 ◽  
Vol 121 (2) ◽  
pp. 309-316 ◽  
Author(s):  
G. A. F. Ritchie ◽  
P. J. Senior ◽  
E. A. Dawes
Keyword(s):  
Substrate Specificity ◽  
Reaction Rate ◽  
Acyl Carrier Protein ◽  
Thiol Group ◽  
Carrier Protein ◽  
Purification And Characterization ◽  
Michaelis Constants ◽  
Coa Reductase ◽  
Acetoacetyl Coa Reductase

A soluble acetoacetyl-CoA reductase (EC 1.1.1.36) was purified 54-fold from Azotobacter beijerinckii N.C.I.B. 9067 and the reaction product identified as d(−)-β-hydroxybutyryl-CoA. The Michaelis constants for acetoacetyl-CoA, NADPH and NADH were determined and the reaction rate was found to be some fivefold greater with NADPH than with NADH. At neutral pH the equilibrium greatly favours the formation of the reduced product. Substrate specificity was in the order: acetoacetyl-CoA>acetoacetylpantetheine>acetoacetyl-(acyl-carrier protein). The enzyme possesses a functional thiol group, suffers inactivation by oxygen and is inhibited by thiol-blocking reagents. Inhibition by p-chloromercuribenzoate is reversed by excess of dithiothreitol, which also protects the enzyme from inactivation by oxygen.

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