Mutation of a basic sequence in the laminin α2LG3 module leads to a lack of proteolytic processing and has different effects on β1 integrin‐mediated cell adhesion and α‐dystroglycan binding

Mutation of a basic sequence in the laminin α2LG3 module leads to a lack of proteolytic processing and has different effects on β1 integrin‐mediated cell adhesion and α‐dystroglycan binding
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层粘连蛋白α2LG3 模块中基本序列的突变导致蛋白水解加工的缺乏,并对β1 整合素介导的细胞粘附和α-肌营养不良聚糖结合产生不同的影响

DOI:
10.1016/s0014-5793(99)01180-1
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发表时间:
1999
期刊:
影响因子:
3.5
通讯作者:
R. Timpl
R. Timpl
中科院分区:
生物学3区
文献类型:
--
作者:
J. Talts;R. Timpl

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在串联阵列α2LG1-3的重组生产过程中,层粘连蛋白α2链的Lg3模块特有的RRKRRQ序列先前被证明对内源性蛋白分解敏感。裂解的多肽键周围的RQ突变并没有阻止这一过程和细胞内的降解。然而,三个交替的碱性残基的丙氨酸突变被证明阻止了α2LG1-3的切割,使α2LG3模块能够获得折叠的球状片段。该突变不改变α2LG1-3的肝素和硫脂结合或细胞黏附,这可能是由α3β1和α6β1整合素介导的。然而,它确实导致了α与营养不良多糖的结合减少了10倍。这些数据支持这样的解释,即肝素/硫脂、β1整合素和α-营养不良糖的结合表位可以占据α2LG1-3结构的不同部分。
A RRKRRQ sequence unique to the LG3 module of the laminin α2 chain was previously shown to be sensitive to endogenous proteolysis during the recombinant production of the tandem array α2LG1-3. Mutation of RQ surrounding the cleaved peptide bond did not prevent this processing and intracellular degradation. Alanine mutagenesis of three alternate basic residues, however, was shown to prevent the cleavage in α2LG1-3, allowing for the α2LG3 module to be obtained as a folded, globular fragment. The mutation did not change heparin and sulfatide binding or cell adhesion of α2LG1-3 which can be mediated by α3β1 and α6β1 integrins. It did, however, cause a 10-fold reduction in α-dystroglycan binding. The data favor the interpretation that binding epitopes for heparin/sulfatides, β1 integrins and α-dystroglycan occupy different parts of the α2LG1-3 structure.
重组层粘连蛋白 G 结构域介导成肌细胞粘附和肝素结合。
DOI: --
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