肌酸激酶同工酶的二乙氨基乙基纤维素色谱法——pH值和血清的影响

DEAE-cellulose chromatography of creatine kinase isoenzymes--effect of pH and serum.

作者信息

Desjardins P R, Rabkin S W, Jacobs H K

出版信息

Clin Biochem. 1979 Jun;12(3):77-82. doi: 10.1016/s0009-9120(79)80070-3.

Abstract

DEAE-cellulose chromatography (pH 7.0) of human heart extracts revealed the presence of three creatine kinase isoenzymes. The CK3 (skeletal muscle) isoenzyme was not retained on the column under these conditions. The CK2 (heart) and CK1 (brain) isoenzymes eluted at a conductivity of 5.5 +/- 0.6 m omega-1 and 11.4 +/- 1.2m omega-1, respectively. When DEAE-cellulose chromatography was performed at pH 8.0, CK2 eluted at a slightly higher conductivity, 6.5 m omega-1, whereas CK1 eluted as before 12.0 m omega-1. DEAE-cellulose chromatography of CK2 and CK1 isoenzymes in the presence of serum protein, and serum albumin had no significant effect on the elution of CK2 at pH 7.0 and 7.4, and on the elution of CK1 at pH 7.0 and 8.0 However, serum and serum albumin decreased the affinity of CK2 for DEAE-celluose at pH 8.0, and caused this isoenzyme to elute at a conductivity of 3.0-3.5 m omega-1. The decreased affinity of CK2 for DEAE-cellulose was not due to aggregation of CK2 with albumin or some other serum protein, but was related to the amount of albumin applied to the column.

摘要

对人心脏提取物进行二乙氨基乙基纤维素色谱分析（pH 7.0），结果显示存在三种肌酸激酶同工酶。在这些条件下，CK3（骨骼肌）同工酶未保留在柱上。CK2（心脏）和CK1（脑）同工酶分别在电导率为5.5±0.6 mΩ⁻¹和11.4±1.2 mΩ⁻¹时洗脱。当在pH 8.0条件下进行二乙氨基乙基纤维素色谱分析时，CK2在稍高的电导率6.5 mΩ⁻¹下洗脱，而CK1仍在12.0 mΩ⁻¹时洗脱。在存在血清蛋白的情况下对CK2和CK1同工酶进行二乙氨基乙基纤维素色谱分析，血清白蛋白对pH 7.0和7.4时CK2的洗脱以及pH 7.0和8.0时CK1的洗脱均无显著影响。然而，血清和血清白蛋白在pH 8.0时降低了CK2对二乙氨基乙基纤维素的亲和力，导致该同工酶在电导率为3.0 - 3.5 mΩ⁻¹时洗脱。CK2对二乙氨基乙基纤维素亲和力的降低并非由于CK2与白蛋白或其他血清蛋白聚集，而是与加到柱上的白蛋白量有关。