Purification and characterization of recombinant mouse and herpes simplex virus ribonucleotide reductase R2 subunit

Biochemistry ◽  
1991 ◽  
Vol 30 (7) ◽  
pp. 1939-1947 ◽  
Author(s):  
Graham J. Mann ◽  
Astrid Graeslund ◽  
El Ichiro Ochiai ◽  
Rolf Ingemarson ◽  
Lars Thelander
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Ribonucleotide Reductase ◽  
Purification And Characterization ◽  
Recombinant Mouse ◽  
Ribonucleotide Reductase R2 ◽  
Simplex Virus ◽  
R2 Subunit

scholarly journals Production of the R2 subunit of ribonucleotide reductase from herpes simplex virus with prokaryotic and eukaryotic expression systems: higher activity of R2 produced by eukaryotic cells related to higher iron-binding capacity

1996 ◽  
Vol 320 (1) ◽  
pp. 129-135 ◽  
Author(s):  
Nathalie LAMARCHE ◽  
Gilles MATTON ◽  
Bernard MASSIE ◽  
Marc FONTECAVE ◽  
Mohamed ATTA ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Ribonucleotide Reductase ◽  
Binding Capacity ◽  
Eukaryotic Expression ◽  
Eukaryotic Cells ◽  
Expression Systems ◽  
Simplex Virus ◽  
R2 Subunit ◽  
Eukaryotic Expression Systems

The R2 subunit of ribonucleotide reductase from herpes simplex virus type 2 was overproduced with prokaryotic and eukaryotic expression systems. The recombinant R2 purified by a two-step procedure exhibited a 3-fold higher activity when produced in eukaryotic cells. Precise quantification of the R2 concentration at each step of the purification indicated that the activity was not altered during the purification procedure. Moreover, we have observed that the level of R2 expression, in eukaryotic cells as well as in prokaryotic cells, did not influence R2 activity. Extensive characterization of the recombinant R2 purified from eukaryotic and prokaryotic expression systems has shown that both types of pure R2 preparations were similar in their 76 kDa dimer contents (more than 95%) and in their ability to bind the R1 subunit. However, we have found that the higher activity of R2 produced in eukaryotic cells is more probably related to a higher capability of binding the iron cofactor as well as a 3-fold greater ability to generate the tyrosyl free radical.

Purification and characterization of proteins excreted by cells infected with herpes simplex virus and their use in diagnosis

Virology ◽  
1978 ◽  
Vol 91 (2) ◽  
pp. 234-242 ◽  
Author(s):  
Anthony B. Chen ◽  
Tamar Ben-Porat ◽  
Richard J. Whitley ◽  
Albert S. Kaplan
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Purification And Characterization ◽  
Simplex Virus
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