[Peroxidase refolding in the presence of specific antibodies].
A panel of eight monoclonal antibodies raised against horseradish root peroxidase has been assembled and characterized. Affinity constants were determined for all antibodies, and their specificity for various structural forms of the enzyme (native peroxidase, apoperoxidase, and denatured peroxidase) were assessed by competitive enzyme immunoassay. The effects of the antibodies on the process of refolding of peroxidase after its denaturing with 6.5 M guanidine hydrochloride were studied spectrophotometrically, by the restoration of the enzymatic activity in the reaction of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonate). The yield of the active enzyme in the course of the refolding was increased 1.5 to 1.7 times in the presence of antibody H1. Effects of the antibodies constituting the panel on the activity of native peroxidase and the stability of its dilute solutions were analyzed.
Bezsudnova
E Iu
EIu
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, 119071 Russia. eubez@inbi.ras.ru
Zherdev
A V
AV
Ermolenko
D N
DN
Iakovleva
I V
IV
Sviridov
V V
VV
Popov
V O
VO
Dzantiev
B B
BB
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D031215:Armoracia / Q000201:enzymology*; D052160:Benzothiazoles; D004795:Enzyme Stability; D019791:Guanidine; D009195:Peroxidase / Q000276:immunology* / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology; D011489:Protein Denaturation; D017510:Protein Folding; D013451:Sulfonic Acids / Q000378:metabolism
D000911:Antibodies, Monoclonal; D052160:Benzothiazoles; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D009195:Peroxidase; D019791:Guanidine