Martín-Martín B, Nabokina S M, Lazo P A, Mollinedo F
Laboratory of Signal Transduction and Leukocyte Biology, Instituto de Biología y Genética Molecular, Facultad de Medicina, Consejo Superior de Investigaciones Científicas-Universidad de Valladolid, Madrid, Spain.
J Leukoc Biol. 1999 Mar;65(3):397-406. doi: 10.1002/jlb.65.3.397.
Syntaxins are major components of vesicle trafficking and their pattern of expression depends on the cell type. Using reverse transcriptase-polymerase chain reaction (RT-PCR), cloning, and sequencing techniques, we have found that human neutrophils and neutrophil-differentiated HL-60 cells co-express syntaxins 1A, 3, 4, 5, 6, 7, 9, 11, and 16. These genes are also expressed in human peripheral blood lymphocytes and SH-SY5Y neuroblastoma cells, which, unlike neutrophils, also expressed syntaxin 10. We have identified two isoforms of syntaxin 3. Syntaxin 3A, similar to the previously reported syntaxin 3, and the novel isoform syntaxin 3B, which is identical to syntaxin 3A but lacks 37 amino acid residues at the carboxy-terminal region. Syntaxin 1 was mainly located to neutrophil granule membranes by confocal microscopy and by immunoblotting of subcellular fractions. These data indicate that syntaxin 1 cannot be considered specific to neural tissues. The level of expression of syntaxins 3, 4, 6, and 11 was increased during neutrophil differentiation of HL-60 cells, whereas that of syntaxins 1A, 5, 9, and 16 was unchanged. Syntaxin 7 was not expressed in undifferentiated HL-60 cells, but its expression was induced on neutrophil differentiation. The expression of several syntaxin genes in human neutrophils could be related to the high secretory capacity of these cells as well as to the presence of different cytoplasmic granules with distinct exocytic capabilities.
syntaxins是囊泡运输的主要组成部分,其表达模式取决于细胞类型。利用逆转录聚合酶链反应(RT-PCR)、克隆和测序技术,我们发现人类中性粒细胞和经中性粒细胞分化的HL-60细胞共表达syntaxins 1A、3、4、5、6、7、9、11和16。这些基因也在人类外周血淋巴细胞和SH-SY5Y神经母细胞瘤细胞中表达,与中性粒细胞不同的是,这些细胞还表达syntaxin 10。我们鉴定出了syntaxin 3的两种异构体。Syntaxin 3A与先前报道的syntaxin 3相似,而新型异构体Syntaxin 3B与Syntaxin 3A相同,但在羧基末端区域缺少37个氨基酸残基。通过共聚焦显微镜和亚细胞组分的免疫印迹分析,Syntaxin 1主要定位于中性粒细胞颗粒膜上。这些数据表明,不能认为Syntaxin 1是神经组织特有的。在HL-60细胞向中性粒细胞分化的过程中,syntaxins 3、4、6和11的表达水平升高,而syntaxins 1A、5、9和16的表达水平没有变化。Syntaxin 7在未分化的HL-60细胞中不表达,但其表达在中性粒细胞分化时被诱导。人类中性粒细胞中几种syntaxin基因的表达可能与这些细胞的高分泌能力以及具有不同胞吐能力的不同细胞质颗粒的存在有关。