人黑色素瘤组织中逆转录酶的生化及免疫学特性

Biochemical and immunological characterization of a reverse transcriptase from human melanoma tissue.

作者信息

Chandra P, Balikcioglu S, Mildner B

出版信息

Cancer Lett. 1978 Dec;5(6):299-310. doi: 10.1016/s0304-3835(78)80055-x.

Abstract

An RNA-direct DNA polymerase was purified from human melanoma tissue by successive column chromatography on DEAE-cellulose (DE-23 and DE-52) and phosphocellulose. The purified reverse transcriptase has a mol. wt. of 68,000, a pH optimum of 8.0, a Mn2+ optimum of 0.6 mM, and a KCl optimum of 60 mM. The purified enzyme transcribes (rA)n - (dT)12, (rC)n - (dG)18, (Ome-rC)n - (dG)18 and a 70s RNA from Rauscher leukemia virus (RLV), but failed to transcribe (dA)n - (dT)12. This enzyme has no terminal deoxynucleotidyl transferase activity. Serological studies have shown that the reverse transcriptase from human melanoma tissue is antigenically not related to DNA polymerases from Simian sarcoma virus (SiSV), Avian myeloblastosis virus (AMV), RLV, and human spleen of a patient with myelofibrosis. The purified enzyme showed a close antigenic resemblance to DNA polymerases from baboon endogenous virus (BEV) and rhabdomyosarcoma virus (RD-114), the endogenous virus of the cat.

摘要

通过在DEAE - 纤维素（DE - 23和DE - 52）和磷酸纤维素上连续柱层析，从人黑色素瘤组织中纯化出一种RNA指导的DNA聚合酶。纯化得到的逆转录酶分子量为68,000，最适pH为8.0，最适Mn²⁺浓度为0.6 mM，最适KCl浓度为60 mM。该纯化酶能转录（rA）n - （dT）12、（rC）n - （dG）18、（Ome - rC）n - （dG）18以及劳斯氏白血病病毒（RLV）的70s RNA，但不能转录（dA）n - （dT）12。这种酶没有末端脱氧核苷酸转移酶活性。血清学研究表明，人黑色素瘤组织中的逆转录酶在抗原性上与猿猴肉瘤病毒（SiSV）、禽成髓细胞瘤病毒（AMV）、RLV以及骨髓纤维化患者人脾脏中的DNA聚合酶无关。纯化后的酶在抗原性上与狒狒内源性病毒（BEV）和猫内源性病毒横纹肌肉瘤病毒（RD - 114）的DNA聚合酶极为相似。