Sequence-activity mapping via depletion reveals striking mutational tolerance and elucidates functional motifs in Tur1a antimicrobial peptide.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
Jonathan Collins, Benjamin J Hackel
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引用次数: 0

Abstract

Proline-rich antimicrobial peptides (PrAMPs) are attractive antibiotic candidates that target gram-negative bacteria ribosomes. We elucidated the sequence-function landscape of 43 000 variants of a recently discovered family member, Tur1a, using the validated SAMP-Dep platform that measures intracellular AMP potency in a high-throughput manner via self-depletion of the cellular host. The platform exhibited high replicate reproducibility (ρ = 0.81) and correlation between synonymous genetic variants (R2 = 0.93). Only two segments within Tur1a exhibited stringent mutational requirements to sustain potency: residues 9YLP11 and 19FP20. This includes the aromatic residue in the hypothesized binding domain but not the PRP domain. Along with unexpected mutational tolerance of PRP, the data contrast hypothesized importance of the 1RRIR4 motif and arginines in general. In addition to mutational tolerance of residue segments with presumed significance, 77% of mutations are functionally neutral. Multimutant performance mainly shows compounding effects from removed combinations of prolines and arginines in addition to the two segments of residues showing individual importance. Several variants identified as active from SAMP-Dep were externally produced and maintained activity when applied to susceptible species exogenously.

通过损耗绘制的序列-活性图揭示了惊人的突变耐受性,并阐明了 Tur1a 抗菌肽中的功能基团。
富脯氨酸抗菌肽(PrAMPs)是针对革兰氏阴性细菌核糖体的有吸引力的候选抗生素。我们利用经过验证的 SAMP-Dep 平台阐明了最近发现的 Tur1a 家族成员的 43000 个变体的序列功能图谱,该平台通过细胞宿主的自我耗竭以高通量方式测量细胞内 AMP 的效力。该平台具有很高的重复再现性(ρ = 0.81)和同义遗传变异之间的相关性(R2 = 0.93)。Tur1a 中只有两个区段显示出维持效力的严格突变要求:残基 9YLP11 和 19FP20。这包括假定结合结构域中的芳香残基,但不包括 PRP 结构域。除了 PRP 的突变耐受性出乎意料之外,数据还对比了 1RRIR4 矩阵和一般精氨酸的假设重要性。除了具有假定重要性的残基段的突变耐受性外,77% 的突变在功能上是中性的。多突变表现主要显示了除显示出单独重要性的两段残基外,去除的脯氨酸和精氨酸组合所产生的复合效应。从 SAMP-Dep 中鉴定出的几种具有活性的变异体是外源生产的,当外源应用于易感物种时仍保持活性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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