Isolation, characterization and partial sequence of cyanogen bromide fragments and thiol peptides from pig kidney d-Amino-acid oxidase

1976 ◽  
Vol 427 (2) ◽  
pp. 634-643 ◽  
Author(s):  
Severino Ronchi ◽  
Lorenzo Minchiotti ◽  
Bruno Curti ◽  
M. Carla Zapponi ◽  
John Bridgen
Keyword(s):  
Amino Acid ◽  
Cyanogen Bromide ◽  
Partial Sequence ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Pig Kidney ◽  
Thiol Peptides

scholarly journals Studies on the active centre of Rhodotorula gracilisd-amino acid oxidase and comparison with pig kidney enzyme

1992 ◽  
Vol 286 (2) ◽  
pp. 389-394 ◽  
Author(s):  
L Pollegioni ◽  
S Ghisla ◽  
M S Pilone
Keyword(s):  
Amino Acid ◽  
Solvent Accessibility ◽  
Alkylating Agents ◽  
Fine Tuning ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Active Centre ◽  
Pig Kidney ◽  
Mammalian Enzyme ◽  
Active Centres

D-Amino acid oxidase (EC 1.4.3.3) from Rhodotorula gracilis has been reconstituted with 8-chloro-, 8-mercapto-, 6-hydroxy-, 2-thio-, 5-deaza- and 1-deaza-FAD, and the properties of the resulting complexes have been studied and compared with those of the correspondingly modified pig kidney D-amino acid oxidases. Binding appears to be tight for most analogues, at least as tight as for native FAD (approximately 10(-8) M). 8-Mercapto- and 6-hydroxy-FAD bind in their para- and ortho-quinoid forms respectively to yeast D-amino acid oxidase, inferring the presence of a positive charge near the flavin N(1) position, as in the case of the mammalian enzyme. On the other hand, important differences in active-site microenvironment emerge: solvent accessibility to flavin position 8 is drastically restricted in yeast D-amino acid oxidase as indicated by the unreactivity of 8-chloro- and 8-mercapto-FAD enzyme with thiolates and alkylating agents. Significantly different microenvironments are also likely to occur around the flavin positions N(1)-C(2) = 0, N(3)-H and N(5). This is deduced from the differences in interaction of the two proteins with 1-deaza-FAD, 5-deaza-FAD and 2-thio-FAD and from the properties of the respective complexes. The same re-side flavin stereospecificity as shown by the mammalian enzyme was determined for the yeast enzyme using 8-hydroxy-5-deaza-FAD. Thus we can deduce the presence of a similar pattern of functional groups at the active centres of the two enzymes, while the fine tuning of specificity and regulation correlate with environmental differences at specific flavin loci.

Improved purification, amino acid analysis and molecular weight of homogeneous d-amino acid oxidase from pig kidney

1973 ◽  
Vol 327 (2) ◽  
pp. 266-273 ◽  
Author(s):  
Bruno Curti ◽  
Severino Ronchi ◽  
Umberto Branzoli ◽  
Giuseppina Ferri ◽  
Charles H. Williams
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Analysis ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Pig Kidney
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