Kawasaki H, Carrera C J, Carson D A
Department of Medicine, University of California, San Diego, La Jolla 92093.
Anal Biochem. 1992 Nov 15;207(1):193-6. doi: 10.1016/0003-2697(92)90522-9.
Deoxycytidine kinase (dCK) is necessary for the activity of several nucleosides used for the chemotherapy of cancer and AIDS. However, the measurement of dCK catalytic activity in crude cell extracts may be imprecise, due to the presence of phosphatases and nucleotidases that degrade the enzyme products. We describe a simple immunoassay for dCK that can measure accurately as little as 5 ng enzyme protein in crude tissue extracts. The assay enabled us to show (i) that mutant cells deficient in dCK activity lack immunoreactive dCK protein, (ii) that dCK catalytic activity and immunoreactivity correlate closely in human tumors, and (iii) that immunoreactive dCK is particularly high in lymphocytes and lymphoid malignancies, although certain solid tumors may also contain the enzyme. The immunoassay of dCK could prove useful in the selection and monitoring of patients who are being treated with nucleosides that are activated by this enzyme.
脱氧胞苷激酶（dCK）对于几种用于癌症和艾滋病化疗的核苷的活性至关重要。然而，由于存在会降解酶产物的磷酸酶和核苷酸酶，在粗细胞提取物中测量dCK催化活性可能不准确。我们描述了一种用于dCK的简单免疫测定法，它能够在粗组织提取物中准确测量低至5 ng的酶蛋白。该测定法使我们能够证明：（i）缺乏dCK活性的突变细胞缺乏免疫反应性dCK蛋白；（ii）在人类肿瘤中，dCK催化活性与免疫反应性密切相关；（iii）免疫反应性dCK在淋巴细胞和淋巴系统恶性肿瘤中特别高，尽管某些实体瘤也可能含有该酶。dCK的免疫测定法可能在选择和监测接受由该酶激活的核苷治疗的患者方面有用。
