Novel γ-sarcoglycan interactors in murine muscle membranes.
Novel γ-sarcoglycan interactors in murine muscle membranes.
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DOI：
10.1186/s13395-021-00285-2
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发表时间：
2022-01-22
期刊：
影响因子：
4.9
通讯作者：
Luna EJ
中科院分区：
文献类型：
--
作者：
Smith TC;Vasilakos G;Shaffer SA;Puglise JM;Chou CH;Barton ER;Luna EJ
肌聚糖复合物（SC）是横纹肌细胞骨架与基底膜连接网络的一部分。SC在机械变形过程中协调磷酸化和Ca++通量的变化，这些过程被γ -肌聚糖（Sgcg）的功能缺失突变所破坏，从而导致肢带肌营养不良2C/R5。为了深入了解SC如何介导肌肉中的机械信号传导，我们利用LC-MS/MS蛋白质组学对富集的肌层组分免疫沉淀物中的SC相关蛋白进行了分析。纳入标准是与来自C57BL/6对照肌的抗sgcg共免疫沉淀，以及在无sgcg肌肉和非特异性IgG的平行实验中代表性不足。在人RH30横纹肌肉瘤细胞共表达实验中验证了相互作用。我们确定了19个候选物作为Sgcg的直接或间接相互作用物，包括其他3个SC蛋白。新的潜在相互作用包括蛋白磷酸酶-1催化亚基- β (Ppp1cb, PP1b)和Na+-K+-Cl -共转运体NKCC1 （SLC12A2）。NKCC1在人RH30横纹肌肉瘤细胞中共表达后与Sgcg共定位，其胞质结构域在免疫沉淀时从细胞溶解物中去除Sgcg，在洗涤剂渗透后与Sgcg共定位。NKCC1在动物体内定位于肌营养不良蛋白复合体附近。布美他尼在孤立肌肉中抑制NKCC1共转运蛋白活性可降低sc依赖性，菌株诱导的细胞外信号调节激酶1和2 （ERK1/2）磷酸化的增加。计算机分析表明，候选SC相互作用物可能与生存信号通路串扰，包括p53、雌激素受体和TRIM25。结果支持NKCC1是一种新的sc相关信号蛋白。此外，其他候选SC相互作用物的特性表明SC和NKCC1以及其他Sgcg相互作用物（如膜-细胞骨架连接物arch绒毛蛋白）可能调节骨骼肌中激酶和Ca++介导的存活信号。在线版本包含补充材料，可在10.1186/s13395-021-00285-2获得。
The sarcoglycan complex (SC) is part of a network that links the striated muscle cytoskeleton to the basal lamina across the sarcolemma. The SC coordinates changes in phosphorylation and Ca++-flux during mechanical deformation, and these processes are disrupted with loss-of-function mutations in gamma-sarcoglycan (Sgcg) that cause Limb girdle muscular dystrophy 2C/R5. To gain insight into how the SC mediates mechano-signaling in muscle, we utilized LC-MS/MS proteomics of SC-associated proteins in immunoprecipitates from enriched sarcolemmal fractions. Criteria for inclusion were co-immunoprecipitation with anti-Sgcg from C57BL/6 control muscle and under-representation in parallel experiments with Sgcg-null muscle and with non-specific IgG. Validation of interaction was performed in co-expression experiments in human RH30 rhabdomyosarcoma cells. We identified 19 candidates as direct or indirect interactors for Sgcg, including the other 3 SC proteins. Novel potential interactors included protein-phosphatase-1-catalytic-subunit-beta (Ppp1cb, PP1b) and Na+-K+-Cl−-co-transporter NKCC1 (SLC12A2). NKCC1 co-localized with Sgcg after co-expression in human RH30 rhabdomyosarcoma cells, and its cytosolic domains depleted Sgcg from cell lysates upon immunoprecipitation and co-localized with Sgcg after detergent permeabilization. NKCC1 localized in proximity to the dystrophin complex at costameres in vivo. Bumetanide inhibition of NKCC1 cotransporter activity in isolated muscles reduced SC-dependent, strain-induced increases in phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2). In silico analysis suggests that candidate SC interactors may cross-talk with survival signaling pathways, including p53, estrogen receptor, and TRIM25. Results support that NKCC1 is a new SC-associated signaling protein. Moreover, the identities of other candidate SC interactors suggest ways by which the SC and NKCC1, along with other Sgcg interactors such as the membrane-cytoskeleton linker archvillin, may regulate kinase- and Ca++-mediated survival signaling in skeletal muscle. The online version contains supplementary material available at 10.1186/s13395-021-00285-2.
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影响因子：
11.4
作者：
Bueno, OF;De Windt, LJ;Molkentin, JD
通讯作者：
Molkentin, JD
DOI：
10.1083/jcb.122.4.809
发表时间：
1993-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ervasti JM;Campbell KP
通讯作者：
Campbell KP
影响因子：
3.3
作者：
Dowling P;Gargan S;Murphy S;Zweyer M;Sabir H;Swandulla D;Ohlendieck K
通讯作者：
Ohlendieck K
影响因子：
4.1
作者：
Cong D;Zhu W;Kuo JS;Hu S;Sun D
通讯作者：
Sun D
影响因子：
4.8
作者：
Bannert, N;Vollhardt, K;Kurth, R
通讯作者：
Kurth, R