Yao Sun, Chunhui Yang, Xin Sun, Yao Li, Qiong Wu, Di Fu, Lei Wang
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A novel approach for the in vitro assay of phospholipase C and its application in analyzing the effect of calcium and pH on populus PsnPLC.
Purpose: Phosphoinositide-specific phospholipase C plays a crucial role in the trans-membrane and lipid signal transduction. Previous studies have elucidated the biological functions of PI-PLC. However, its function mechanism has not been fully understood.
Results: We present a novel approach to analyze the in vitro activity of phospholipase C from Populus simonii x P. nigra. The results indicate that the optimal condition for PsnPLC activity is pH 7.4/10 μM Ca2+. The increase of Ca2+ and pH, as well as the deletion of the EF-hand domain or C2 domain, will lead to the reduced PsnPLC activity. Electrophoresis migration shift assays confirm that PsnPLC could bind Ca2+ in vitro, while conditions of Ca2+ binding and pH 8.8 might lead to its degradation.
Conclusion: The findings identify Ca2+ and pH as key factors modulating PsnPLC function, which will provide evidence linking PI-PLC and the Ca2+ signaling network.
期刊介绍：
Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them.
All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included.
Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields.
The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories.
Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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