In vitro evolution driven by epistasis reveals alternative cholesterol-specific binding motifs of perfringolysin O

被引：1

作者：

Sakanovic, Aleksandra ^{[1
]}

Kranjc, Nace ^{[1
]}

Omersa, Neza ^{[1
]}

Aden, Sasa ^{[1
]}

Kezar, Andreja ^{[1
]}

Kisovec, Matic ^{[1
]}

Zavec, Apolonija Bedina ^{[1
]}

Caserman, Simon ^{[1
]}

Gilbert, Robert J. C. ^{[2
]}

Podobnik, Marjetka ^{[1
]}

Crnkovic, Ana ^{[1
]}

Anderluh, Gregor ^{[1
]}

机构：

[1] Natl Inst Chem, Dept Mol Biol & Nanobiotechnol, Ljubljana, Slovenia

[2] Univ Oxford, Wellcome Ctr Human Genet, Div Struct Biol, Oxford, England

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2024年 / 300卷 / 09期

关键词：

THIOL-ACTIVATED CYTOLYSIN; DEPENDENT CYTOLYSINS; PORE FORMATION; PROTEIN; SEQUENCE; TOXIN; PNEUMOLYSIN; INSIGHTS; AFFINITY; REQUIRE;

D O I：

10.1016/j.jbc.2024.107664

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The crucial molecular factors that shape the interfaces of lipid-binding proteins with their target ligands and surfaces remain unknown due to the complex makeup of biological membranes. Cholesterol, the major modulator of bilayer structure in mammalian cell membranes, is recognized by various proteins, including the well-studied cholesteroldependent cytolysins. Here, we use in vitro evolution to investigate the molecular adaptations that preserve the cholesterol specificity of perfringolysin O, the prototypical cholesterol-dependent cytolysin from Clostridium perfringens. We identify variants with altered membrane-binding interfaces whose cholesterol-specific activity exceeds that of the wild-type perfringolysin O. These novel variants represent alternative evolutionary outcomes and have mutations at conserved positions that can only accumulate when epistatic constraints are alleviated. Our results improve the current understanding of the biochemical malleability of the surface of a lipid-binding protein.

引用

页数：16