HUMAN MYELOID CHEMOTACTIC RECEPTOR: STRUCTURE-FUNCTION

本提案探讨了生化结构、代谢 人外周血单个核细胞表面甲酰基肽趋化性受体及其功能 外周血白细胞 这是一项长期研究的开始， 人骨髓膜发育和功能。 这个受体， 天然的或共价亲和标记的，将从人 中性粒细胞或培养的骨髓细胞系（HL-60和U937），通过 经典和亲和技术，并根据 蛋白酶结构域和肽图谱、疏水性和分子大小。 会产生抗受体抗体，并自发产生抗受体抗体 将在抗粒细胞抗体患者的血清中寻找抗体， 抗体的 受体介导配体的重要性及其作用机制 聚集和内吞作用将使用 荧光标记的激动剂和拮抗剂，多价拮抗剂 和抗受体抗体。 受体在细胞内的分布 膜和邻近的膜蛋白将被探索。 率 受体的合成和周转将在髓样细胞中确定， 使用重同位素氨基酸标记和CsCl 2梯度的成熟 离心法 趋化性受体的结构和加工将是 在异常骨髓发育（白血病， 骨髓增生异常疾病，阵发性睡眠性血红蛋白尿，骨 骨髓移植或细胞毒性化疗）、细菌性败血症或 复发性细菌感染和使用抗炎药物期间。 两 涉及科学学科。 血液学，因为这个提议 研究血白细胞功能，HL-60分化是一个模型， 骨髓和几种血液学中白细胞发育系统 疾病将被研究。 免疫学，因为受体介导 免疫重要的白细胞反应。

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.