Interaction of Fc gamma receptor type IIIB with complement receptor type 3 in fibroblast transfectants: evidence from lateral diffusion and resonance energy transfer studies.
Interaction of Fc gamma receptor type IIIB with complement receptor type 3 in fibroblast transfectants: evidence from lateral diffusion and resonance energy transfer studies.
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成纤维细胞转染子中 Fc γ 受体 IIIB 型与补体受体 3 型的相互作用：来自横向扩散和共振能量转移研究的证据。
DOI：
10.1006/jmbi.1995.0166
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Petty,HR
中科院分区：
文献类型：
--
作者：
Poo,H;Krauss,JC;Mayo-Bond,L;Todd3rd,RF;Petty,HR
为了探索FcγRIIIB（一种GPI连接蛋白）与白细胞整联蛋白CR 3之间的潜在受体间相互作用，我们制备了表达FcγRIIIB、CR 3或FcγRIIIB和CR 3的转染3 T3成纤维细胞系。我们使用光漂白后荧光恢复（FRAP）和共振能量转移（r.e.t.）来检验FcγRIIIB和CR 3在膜中物理缔合的假设。显微镜单独表达FcγRIIIB的细胞显示出3.4×10−9（±2.9×10−9）cm 2/s的扩散系数（D）和移动的分数（m.f.）0.73（±0.10）。相比之下，FcγRIIIB显示D = 2.5×10−9（±1.4×10−9）cm 2/秒（n.s.）和一个m.f. 0.48（±0.08）（p < 0.01），因此表明CR 3的共表达抑制了FcγRIIIB的侧向扩散。为了进一步测试这些基因产物之间的直接物理相互作用，（r.e.t.）进行显微镜检查。表达两种受体的细胞上的供体标记抗CR 3和受体标记抗Fc γRIIIB产生r.e.t.光子计数率为8.9（±6.4）千计数/秒（kC/s），而CR 3对CR 3测量值为1.6（±0.6）kC/s（p < 0.01）。此外，添加外源性试剂，如N-乙酰基-D-葡糖胺，但不是吲哚美辛，减少了这些相互作用的转染膜的大小。这些数据支持FcγRIIIB亚群与CR 3物理相关的观点，并且这种相关性可能受到外源性化合物的影响。
To explore potential inter-receptor interactions between FcγRIIIB, a GPI-linked protein, and the leukocyte integrin CR3, we have prepared transfected 3T3 fibroblast cell lines expressing FcγRIIIB, CR3, or both FcγRIIIB and CR3. We test the hypothesis that FcγRIIIB and CR3 are physically associated in membranes using fluorescence recovery after photobleaching (FRAP) and resonance energy transfer (r.e.t.) microscopy. Cells expressing FcγRIIIB alone displayed a diffusion coefficient (D) of 3.4×10−9(±2.9×10−9) cm2/second and a mobile fraction (m.f.) of 0.73 (±0.10). In contrast, FcγRIIIB exhibited D = 2.5×10−9(±1.4×10−9) cm2/second (n.s.) and a m.f. of 0.48 (±0.08) (p < 0.01) on cells expressing both FcγRIIB and CR3, thus indicating that co-expression of CR3 constrains the lateral diffusion of FcγRIIIB. To further test for a direct physical interaction between these gene products, (r.e.t.) microscopy was performed. Donor-labeled anti-CR3 and acceptor-labeled anti-FcγRIIIB on cells expressing both receptors yielded a r.e.t. photon count rate of 8.9(±6.4) kilocounts/second (kC/s), whereas CR3-to-CR3 measurements gave 1.6(±0.6) kC/s (p < 0.01). Moreover, the addition of exogenous agents such as N-acetyl-d-glucosamine, but not indomethacin, diminished the magnitude of these interactions in transfectant membranes. These data support the notion that a subpopulation of FcγRIIIB is physically associated with CR3 and that this association can be affected by exogeneous compounds.