8GL3

De novo design of monomeric helical bundles for pH-controlled membrane lysis

Summary for 8GL3

• Entry DOI: 10.2210/pdb8gl3/pdb
• Descriptor: pRLB-519 (2 entities in total)
• Functional Keywords: protein design, ph-responsive, membrane lysis, de novo protein
• Biological source: synthetic construct
• Total number of polymer chains: 1
• Total formula weight: 27599.22

Authors

Goldbach, N.,Bera, A.K.,Baker, D.,Kang, A. (deposition date: 2023-03-20, release date: 2023-09-06, Last modification date: 2023-11-08)

Primary citation

Goldbach, N.,Benna, I.,Wicky, B.I.M.,Croft, J.T.,Carter, L.,Bera, A.K.,Nguyen, H.,Kang, A.,Sankaran, B.,Yang, E.C.,Lee, K.K.,Baker, D.
De novo design of monomeric helical bundles for pH-controlled membrane lysis.
Protein Sci., 32:e4769-e4769, 2023

PubMed Abstract: Targeted intracellular delivery via receptor-mediated endocytosis requires the delivered cargo to escape the endosome to prevent lysosomal degradation. This can in principle be achieved by membrane lysis tightly restricted to endosomal membranes upon internalization to avoid general membrane insertion and lysis. Here, we describe the design of small monomeric proteins with buried histidine containing pH-responsive hydrogen bond networks and membrane permeating amphipathic helices. Of the 30 designs that were experimentally tested, all expressed in Escherichia coli, 13 were monomeric with the expected secondary structure, and 4 designs disrupted artificial liposomes in a pH-dependent manner. Mutational analysis showed that the buried histidine hydrogen bond networks mediate pH-responsiveness and control lysis of model membranes within a very narrow range of pH (6.0-5.5) with almost no lysis occurring at neutral pH. These tightly controlled lytic monomers could help mediate endosomal escape in designed targeted delivery platforms.

PubMed: 37632837
DOI: 10.1002/pro.4769

Experimental method

X-RAY DIFFRACTION (2.3 Å)