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抑制细胞集落和簇形成的白血病相关抑制活性的分离与生化特性分析

Isolation and biochemical characterization of leukemia-associated inhibitory activity that suppresses colony and cluster formation of cells.

作者信息

Bognacki J, Broxmeyer H E, Lobue J

出版信息

Biochim Biophys Acta. 1981 Jan 21;672(2):176-90. doi: 10.1016/0304-4165(81)90391-3.

Abstract

Leukemia-associated inhibitory activity suppresses colony and cluster formation in vitro cells derived from granulocyte-macrophage progenitor cells of normal donors. It does not inhibit these same progenitor cells from patients with leukemia and it may contribute to the proliferative advantage leukemia cells appear to possess over normal hematopoietic cells during acute leukemia. The inhibitory activity was isolated by a combination of procedures including: ultracentrifugation, Sephadex G-200, carboxymethylcellulose, SDS-polyacrylamide gel electrophoresis, thin-layer and preparative isoelectric focusing and concanavalin A-Sepharose. Leukemia-associated inhibitory activity was characterized as a glycoprotein. it was inactivated by trypsin, chymotrypsin, pronase and periodate treatment. It bound to and was eluted by alpha-methylmannose from concanavalin A-Sepharose columns and had an apparent Mr range of 450-550 000 and an isoelectric focus value between pH 4.6 and 4.9. Crude leukemia associated inhibitory activity was temperature sensitive but the more purified preparations were heat stable.

摘要

白血病相关抑制活性可抑制来自正常供体粒细胞-巨噬细胞祖细胞的体外细胞形成集落和集簇。它并不抑制白血病患者的这些相同祖细胞,并且可能有助于白血病细胞在急性白血病期间似乎比正常造血细胞所具有的增殖优势。通过包括超速离心、葡聚糖凝胶G-200、羧甲基纤维素、SDS-聚丙烯酰胺凝胶电泳、薄层和制备性等电聚焦以及伴刀豆球蛋白A-琼脂糖等一系列程序组合分离出该抑制活性。白血病相关抑制活性被鉴定为一种糖蛋白。它可被胰蛋白酶、糜蛋白酶、链霉蛋白酶和高碘酸盐处理灭活。它与伴刀豆球蛋白A-琼脂糖柱上的α-甲基甘露糖结合并被洗脱,表观分子量范围为450 - 550 000,等电聚焦值在pH 4.6至4.9之间。粗制的白血病相关抑制活性对温度敏感,但纯化程度更高的制剂热稳定。

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