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Transglutaminase-mediated crosslinking of a host defence peptide derived from human apolipoprotein B and its effect on the peptide antimicrobial activity
Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 3 ) Pub Date : 2020-11-27 , DOI: 10.1016/j.bbagen.2020.129803
Eliana Dell'Olmo , Rosa Gaglione , Angela Arciello , Renata Piccoli , Valeria Cafaro , Antimo Di Maro , Sara Ragucci , Raffaele Porta , C. Valeria L. Giosafatto

Background

Microbial transglutaminase (mTG) has been successfully used to produce site-specific protein conjugates derivatized at the level of Gln and/or Lys residues for different biotechnological applications. Here, a recombinant peptide identified in human apolipoprotein B sequence, named r(P)ApoBL and endowed with antimicrobial activity, was studied as a possible acyl acceptor substrate of mTG with at least one of the six Lys residues present in its sequence.

Methods

The enzymatic crosslinking reaction was performed in vitro using N,N-dimethylcasein, substance P and bitter vetch (Vicia ervilia) seed proteins, well known acyl donor substrates in mTG-catalyzed reactions. Mass spectrometry analyses were performed for identifying the Lys residue(s) involved in the crosslinking reaction. Finally, bitter vetch protein-based antimicrobial films grafted with r(P)ApoBL were prepared and, their biological activity evaluated.

Results

r(P)ApoBL was able to be enzymatically modified by mTG. In particular, it was demonstrated the highly selective crosslinking of the peptide under study by mTG at level of Lys-18. Interestingly, the biological activity of the peptide when grafted into protein-based films was found to be lost following mTG-catalyzed crosslinking.

Conclusions

r(P)ApoBL was shown to be an effective acyl acceptor substrate of mTG. The involvement of Lys-18 in the enzymatic reaction was demonstrated. In addition, films grafted with r(P)ApoBL in the presence of mTG lost antimicrobial property.

General significance

A possible role of mTG as biotechnological tool to modulate the r(P)ApoBL antimicrobial activity was hypothesized, and a potential use in food packaging of protein-based films grafted with r(P)ApoBL was suggested.



中文翻译:

转谷氨酰胺酶介导的人载脂蛋白B宿主防御肽的交联及其对肽抗菌活性的影响

背景

微生物转谷氨酰胺酶(mTG)已成功用于生产针对不同生物技术应用在Gln和/或Lys残基水平上衍生化的位点特异性蛋白偶联物。在此,研究了在人类载脂蛋白B序列中鉴定出的重组肽,称为r(P)ApoB L,具有抗菌活性,被认为是mTG的可能的酰基受体底物,在其序列中存在至少六个Lys残基。

方法

酶促交联反应进行体外使用ÑÑ -dimethylcasein,P物质和苦野豌豆(蚕豆ervilia中的mTG催化的反应)的种子蛋白,公知的酰基供体底物。进行质谱分析以鉴定参与交联反应的Lys残基。最后,制备了嫁接了r(P)ApoB L的基于苦etch菜蛋白的抗菌膜,并对其生物学活性进行了评估。

结果

r(P)ApoB L能够被mTG酶促修饰。特别地,通过mTG在Lys-18水平上证明了所研究的肽的高度选择性交联。有趣的是,发现在mTG催化的交联后,该肽的生物活性被嫁接到基于蛋白质的薄膜中。

结论

r(P)ApoB L被证明是mTG的有效酰基受体底物。证明了Lys-18参与酶促反应。此外,在mTG存在下用r(P)ApoB L接枝的薄膜失去了抗菌性能。

一般意义

推测了mTG作为调节r(P)ApoB L抗菌活性的生物技术工具的可能作用,并提出了在食品包装中嫁接了r(P)ApoB L的蛋白质基薄膜的潜在用途。

更新日期:2020-12-07
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