Abstract
Hydrogen peroxide removal activities in normal and acatalasemic mouse hemolysates were examined to determine the optimal temperature of catalase. From thermal stability of the removal activities in hemolysates, the removal activities were divided into two activities. The removal activity deactivated at lower temperature was catalase, and the 50% inactivation was observed after 10 min incubation at 47.2 ± 0.5°C for normal hemolysates and 34.0 ± 0.8°C for acatalasemic ones. The removal activity deactivated at a higher temperature remained after the addition of sodium azide, and the 50% inactivation was observed at 63.5 ± 1.4°C. After separation of the removal activities by carboxymethyl-cellulose column chromatography, the removal activity deactivated at higher temperature was attributed to the activity by hemoglobin. From Lineweaver-Burk plot analysis of the removal rates by hemoglobin at 37°C, the Michaelis constant for hydrogen peroxide and the maximum velocity were 201 ± 53 μM and 5.37 ± 1.39 μmol/s per g of Hb, respectively. Removal rates by hemoglobin in mouse hemolysates at 37°C in 70 μM hydrogen peroxide were 1.32 ± 0.12 μmol/s per g of Hb. Catalase activity (k/g Hb: rate constant related to the hemoglobin content) in normal mouse hemolysates was 104 ± 12 at 25°C and 117 ± 10 at 37°C, and that in acatalasemic hemolysates was 10.5 ± 1.7 at 25°C. These results indicate that activity of hydrogen peroxide removal by hemoglobin is substantial and the activity in acatalasemic hemolysates is predominant at low concentration of hydrogen peroxide.
| Original language | English |
|---|---|
| Pages (from-to) | 131-137 |
| Number of pages | 7 |
| Journal | Biochimica et Biophysica Acta - Molecular Basis of Disease |
| Volume | 1361 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Aug 22 1997 |
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Keywords
- Acatalasemic erythrocyte
- Catalase
- Hemoglobin
- Hydrogen peroxide removal rate
- Mouse hemolysate
- Takahara disease
ASJC Scopus subject areas
- Molecular Biology
- Molecular Medicine
- Biophysics
Cite this
Characterization of hydrogen peroxide removal activities in mouse hemolysates : Catalyse activity and hydrogen peroxide removal activity by hemoglobin. / Masuoka, Noriyoshi; Wakimoto, Masahiro; Ohta, Jun; Ishii, Kunihiko; Nakano, Taku.
In: Biochimica et Biophysica Acta - Molecular Basis of Disease, Vol. 1361, No. 2, 22.08.1997, p. 131-137.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of hydrogen peroxide removal activities in mouse hemolysates
T2 - Catalyse activity and hydrogen peroxide removal activity by hemoglobin
AU - Masuoka, Noriyoshi
AU - Wakimoto, Masahiro
AU - Ohta, Jun
AU - Ishii, Kunihiko
AU - Nakano, Taku
PY - 1997/8/22
Y1 - 1997/8/22
N2 - Hydrogen peroxide removal activities in normal and acatalasemic mouse hemolysates were examined to determine the optimal temperature of catalase. From thermal stability of the removal activities in hemolysates, the removal activities were divided into two activities. The removal activity deactivated at lower temperature was catalase, and the 50% inactivation was observed after 10 min incubation at 47.2 ± 0.5°C for normal hemolysates and 34.0 ± 0.8°C for acatalasemic ones. The removal activity deactivated at a higher temperature remained after the addition of sodium azide, and the 50% inactivation was observed at 63.5 ± 1.4°C. After separation of the removal activities by carboxymethyl-cellulose column chromatography, the removal activity deactivated at higher temperature was attributed to the activity by hemoglobin. From Lineweaver-Burk plot analysis of the removal rates by hemoglobin at 37°C, the Michaelis constant for hydrogen peroxide and the maximum velocity were 201 ± 53 μM and 5.37 ± 1.39 μmol/s per g of Hb, respectively. Removal rates by hemoglobin in mouse hemolysates at 37°C in 70 μM hydrogen peroxide were 1.32 ± 0.12 μmol/s per g of Hb. Catalase activity (k/g Hb: rate constant related to the hemoglobin content) in normal mouse hemolysates was 104 ± 12 at 25°C and 117 ± 10 at 37°C, and that in acatalasemic hemolysates was 10.5 ± 1.7 at 25°C. These results indicate that activity of hydrogen peroxide removal by hemoglobin is substantial and the activity in acatalasemic hemolysates is predominant at low concentration of hydrogen peroxide.
AB - Hydrogen peroxide removal activities in normal and acatalasemic mouse hemolysates were examined to determine the optimal temperature of catalase. From thermal stability of the removal activities in hemolysates, the removal activities were divided into two activities. The removal activity deactivated at lower temperature was catalase, and the 50% inactivation was observed after 10 min incubation at 47.2 ± 0.5°C for normal hemolysates and 34.0 ± 0.8°C for acatalasemic ones. The removal activity deactivated at a higher temperature remained after the addition of sodium azide, and the 50% inactivation was observed at 63.5 ± 1.4°C. After separation of the removal activities by carboxymethyl-cellulose column chromatography, the removal activity deactivated at higher temperature was attributed to the activity by hemoglobin. From Lineweaver-Burk plot analysis of the removal rates by hemoglobin at 37°C, the Michaelis constant for hydrogen peroxide and the maximum velocity were 201 ± 53 μM and 5.37 ± 1.39 μmol/s per g of Hb, respectively. Removal rates by hemoglobin in mouse hemolysates at 37°C in 70 μM hydrogen peroxide were 1.32 ± 0.12 μmol/s per g of Hb. Catalase activity (k/g Hb: rate constant related to the hemoglobin content) in normal mouse hemolysates was 104 ± 12 at 25°C and 117 ± 10 at 37°C, and that in acatalasemic hemolysates was 10.5 ± 1.7 at 25°C. These results indicate that activity of hydrogen peroxide removal by hemoglobin is substantial and the activity in acatalasemic hemolysates is predominant at low concentration of hydrogen peroxide.
KW - Acatalasemic erythrocyte
KW - Catalase
KW - Hemoglobin
KW - Hydrogen peroxide removal rate
KW - Mouse hemolysate
KW - Takahara disease
UR - http://www.scopus.com/inward/record.url?scp=0030854719&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030854719&partnerID=8YFLogxK
U2 - 10.1016/S0925-4439(97)00024-0
DO - 10.1016/S0925-4439(97)00024-0
M3 - Article
VL - 1361
SP - 131
EP - 137
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
SN - 0925-4439
IS - 2
ER -