Structure and Function of the Bacterial Protein Toxin Phenomycin

Bente Kring Hansen; Camilla K Larsen; Jakob T Nielsen; Esben B Svenningsen; Lan B Van; Kristian M Jacobsen; Morten Bjerring; Rasmus K Flygaard; Lasse B Jenner; Lene N Nejsum; Ditlev E Brodersen; Frans A A Mulder; Thomas Tørring; Thomas B. Poulsen

doi:10.1016/j.str.2020.03.003

Structure and Function of the Bacterial Protein Toxin Phenomycin

Bente Kring Hansen, Camilla K Larsen, Jakob T Nielsen, Esben B Svenningsen, Lan B Van, Kristian M Jacobsen, Morten Bjerring, Rasmus K Flygaard, Lasse B Jenner, Lene N Nejsum, Ditlev E Brodersen, Frans A A Mulder, Thomas Tørring, Thomas B. Poulsen

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

2 Citations (Scopus)

Abstract

Phenomycin is a bacterial mini-protein of 89 amino acids discovered more than 50 years ago with toxicity in the nanomolar regime toward mammalian cells. The protein inhibits the function of the eukaryotic ribosome in cell-free systems and appears to target translation initiation. Several fundamental questions concerning the cellular activity of phenomycin, however, have remained unanswered. In this paper, we have used morphological profiling to show that direct inhibition of translation underlies the toxicity of phenomycin in cells. We have performed studies of the cellular uptake mechanism of phenomycin, showing that endosomal escape is the toxicity-limiting step, and we have solved a solution phase high-resolution structure of the protein using NMR spectroscopy. Through bioinformatic as well as functional comparisons between phenomycin and two homologs, we have identified a peptide segment, which constitutes one of two loops in the structure that is critical for the toxicity of phenomycin.

Original language	English
Journal	Structure
Volume	28
Issue	5
Pages (from-to)	528-539.e9
Number of pages	12
ISSN	0969-2126
DOIs	https://doi.org/10.1016/j.str.2020.03.003
Publication status	Published - 2020

Keywords

NMR structure
cell-penetrating peptide
mini-protein
natural product
protein synthesis inhibitor
protein toxin
ribosome

Access to Document

10.1016/j.str.2020.03.003

https://www.biorxiv.org/content/biorxiv/early/2019/11/29/847772.full.pdf

Cite this

@article{b55a6a6830c841f5a29710d15998bede,

title = "Structure and Function of the Bacterial Protein Toxin Phenomycin",

abstract = "Phenomycin is a bacterial mini-protein of 89 amino acids discovered more than 50 years ago with toxicity in the nanomolar regime toward mammalian cells. The protein inhibits the function of the eukaryotic ribosome in cell-free systems and appears to target translation initiation. Several fundamental questions concerning the cellular activity of phenomycin, however, have remained unanswered. In this paper, we have used morphological profiling to show that direct inhibition of translation underlies the toxicity of phenomycin in cells. We have performed studies of the cellular uptake mechanism of phenomycin, showing that endosomal escape is the toxicity-limiting step, and we have solved a solution phase high-resolution structure of the protein using NMR spectroscopy. Through bioinformatic as well as functional comparisons between phenomycin and two homologs, we have identified a peptide segment, which constitutes one of two loops in the structure that is critical for the toxicity of phenomycin.",

keywords = "NMR structure, cell-penetrating peptide, mini-protein, natural product, protein synthesis inhibitor, protein toxin, ribosome",

author = "Hansen, {Bente Kring} and Larsen, {Camilla K} and Nielsen, {Jakob T} and Svenningsen, {Esben B} and Van, {Lan B} and Jacobsen, {Kristian M} and Morten Bjerring and Flygaard, {Rasmus K} and Jenner, {Lasse B} and Nejsum, {Lene N} and Brodersen, {Ditlev E} and Mulder, {Frans A A} and Thomas T{\o}rring and Poulsen, {Thomas B.}",

year = "2020",

doi = "10.1016/j.str.2020.03.003",

language = "English",

volume = "28",

pages = "528--539.e9",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "5",

}

TY - JOUR

T1 - Structure and Function of the Bacterial Protein Toxin Phenomycin

AU - Hansen, Bente Kring

AU - Larsen, Camilla K

AU - Nielsen, Jakob T

AU - Svenningsen, Esben B

AU - Van, Lan B

AU - Jacobsen, Kristian M

AU - Bjerring, Morten

AU - Flygaard, Rasmus K

AU - Jenner, Lasse B

AU - Nejsum, Lene N

AU - Brodersen, Ditlev E

AU - Mulder, Frans A A

AU - Tørring, Thomas

AU - Poulsen, Thomas B.

PY - 2020

Y1 - 2020

N2 - Phenomycin is a bacterial mini-protein of 89 amino acids discovered more than 50 years ago with toxicity in the nanomolar regime toward mammalian cells. The protein inhibits the function of the eukaryotic ribosome in cell-free systems and appears to target translation initiation. Several fundamental questions concerning the cellular activity of phenomycin, however, have remained unanswered. In this paper, we have used morphological profiling to show that direct inhibition of translation underlies the toxicity of phenomycin in cells. We have performed studies of the cellular uptake mechanism of phenomycin, showing that endosomal escape is the toxicity-limiting step, and we have solved a solution phase high-resolution structure of the protein using NMR spectroscopy. Through bioinformatic as well as functional comparisons between phenomycin and two homologs, we have identified a peptide segment, which constitutes one of two loops in the structure that is critical for the toxicity of phenomycin.

AB - Phenomycin is a bacterial mini-protein of 89 amino acids discovered more than 50 years ago with toxicity in the nanomolar regime toward mammalian cells. The protein inhibits the function of the eukaryotic ribosome in cell-free systems and appears to target translation initiation. Several fundamental questions concerning the cellular activity of phenomycin, however, have remained unanswered. In this paper, we have used morphological profiling to show that direct inhibition of translation underlies the toxicity of phenomycin in cells. We have performed studies of the cellular uptake mechanism of phenomycin, showing that endosomal escape is the toxicity-limiting step, and we have solved a solution phase high-resolution structure of the protein using NMR spectroscopy. Through bioinformatic as well as functional comparisons between phenomycin and two homologs, we have identified a peptide segment, which constitutes one of two loops in the structure that is critical for the toxicity of phenomycin.

KW - NMR structure

KW - cell-penetrating peptide

KW - mini-protein

KW - natural product

KW - protein synthesis inhibitor

KW - protein toxin

KW - ribosome

UR - http://www.scopus.com/inward/record.url?scp=85083019610&partnerID=8YFLogxK

U2 - 10.1016/j.str.2020.03.003

DO - 10.1016/j.str.2020.03.003

M3 - Journal article

C2 - 32220302

SN - 0969-2126

VL - 28

SP - 528-539.e9

JO - Structure

JF - Structure

IS - 5

ER -

Structure and Function of the Bacterial Protein Toxin Phenomycin

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this