HUMAN MYELOID CHEMOTACTIC RECEPTOR: STRUCTURE-FUNCTION

这项建议探讨了生化结构、新陈代谢的几个方面 存在于人体的甲酰肽趋化受体及其功能 外周血白细胞。这是一个长期研究的开始 人髓系膜的发育和功能。这种受体要么是 天然的或共价亲和标记的，将从人类中溶解 中性粒细胞或培养的髓系细胞系(HL-60和U937)，用 经典和亲和技术，其特征在于 蛋白酶域和肽图、疏水性和分子大小。 会产生抗受体抗体和自发的抗受体抗体 将在抗粒细胞患者的血清中寻找抗体 抗体。受体介导的配体的重要性及其作用机制 将使用以下方法研究聚集和内吞作用 荧光标记的激动剂和抗激动剂、多价拮抗剂 和抗受体抗体。受体在脑内的分布 膜和邻近的膜蛋白将被探索。这一速度 受体的合成和周转将在髓样细胞期间确定 重同位素氨基酸标记和CsCl2梯度成熟法 离心法。趋化受体的结构和加工将是 研究对象为骨髓发育异常的患者(白血病、 骨后性骨髓增生性疾病，阵发性睡眠性血红蛋白尿 骨髓移植或细胞毒性化疗)、细菌性败血症或 反复的细菌感染和抗炎药物的使用。二 科学学科也牵涉其中。血液学，因为提案 处理血白细胞功能，HL-60分化为模型 骨髓和几种血液学中的白细胞发育系统 将对疾病进行研究。和免疫学，因为受体介导 免疫上重要的白细胞反应。

This proposal explores several aspects of biochemical structure, metabolism and function of the formyl peptide chemotactic receptor present on human peripheral blood leukocytes. This is the beginning of a long-term study of human myeloid membrane development and function. This receptor, either native or covalently affinity-labeled, will be solubilized from human neutrophils or cultured myeloid cell lines (HL-60 and U937), purified by classical and affinity techniques and characterized with respect to protease domains and peptide maps, hydrophobicity, and molecular size. Anti-receptor antibodies will be produced and spontaneous anti-receptor antibodies will be sought in serum from patients with anti-granulocyte antibodies. The importance and mechanism of receptor-mediated ligand aggregation and endocytosis will be investigated using fluorescently-labeled agonists and antagoinists, multivalent antagonists and anti-receptor antibodies. The disposition of the receptor within the membrane and neighboring membrane proteins will be explored. The rate of receptor synthesis and turnover will be determined during myeloid maturation using heavy isotope amino acid labeling and CsCl2 gradient centrifugation. Chemotactic receptor structure and processing will be studied in patients with abnormal myeloid development (leukemias, myelodysplastic diseases, paroxysmal nocturnal hemoglobinuria, after bone marrow transplantation or cytotoxic chemotherapy), bacterial sepsis or recurrent bacterial infection and during anti-inflammatory drug use. Two scientific disciplines are involved. Hematology, because the proposal deals with blood leukocyte function, HL-60 differentiation is a model system for leukocyte development in the bone marrow and several hematologic diseases will be studied. And Immunology, because the receptor mediates immunologically important leukocyte responses.