Proprotein convertases generate a highly functional heterodimeric form of thymic stromal lymphopoietin in humans

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Abstract

Rationale Thymic stromal lymphopoietin (TSLP) is known to be elevated and truncated in nasal polyps (NPs) of patients with chronic rhinosinusitis and might play a significant role in type 2 inflammation in this disease. However, neither the structure nor the role of the truncated products of TSLP has been studied. Objective We sought to investigate the mechanisms of truncation of TSLP in NPs and the function of the truncated products. Methods We incubated recombinant human TSLP with NP extracts, and determined the protein sequence of the truncated forms of TSLP using Edman protein sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. We investigated the functional activity of truncated TSLP using a PBMC-based bioassay. Results Edman sequencing and mass spectrometry results indicated that NP extracts generated 2 major truncated products, TSLP (residues 29-124) and TSLP (131-159). Interestingly, these 2 products remained linked with disulfide bonds and presented as a dimerized form, TSLP (29-124 + 131-159). We identified that members of the proprotein convertase were rate-limiting enzymes in the truncation of TSLP between residues 130 and 131 and generated a heterodimeric unstable metabolite TSLP (29-130 + 131-159). Carboxypeptidase N immediately digested 6 amino acids from the C terminus of the longer subunit of TSLP to generate a stable dimerized form, TSLP (29-124 + 131-159), in NPs. These truncations were homeostatic but primate-specific events. A metabolite TSLP (29-130 + 131-159) strongly activated myeloid dendritic cells and group 2 innate lymphoid cells compared with mature TSLP. Conclusions Posttranslational modifications control the functional activity of TSLP in humans and overproduction of TSLP may be a key trigger for the amplification of type 2 inflammation in diseases.

Original languageEnglish (US)
Pages (from-to)1559-1567.e8
JournalJournal of Allergy and Clinical Immunology
Volume139
Issue number5
DOIs
StatePublished - May 1 2017

Fingerprint

Proprotein Convertases
Nasal Polyps
thymic stromal lymphopoietin
Mass Spectrometry
Lysine Carboxypeptidase
Inflammation

Keywords

  • Carboxypeptidase
  • chronic rhinosinusitis
  • dendritic cells
  • group 2 innate lymphoid cells
  • nasal polyps
  • posttranslational modification
  • proprotein convertases
  • thymic stromal lymphopoietin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

@article{8054cf67c29a436db2ba871df8f0cc9e,
title = "Proprotein convertases generate a highly functional heterodimeric form of thymic stromal lymphopoietin in humans",
abstract = "Rationale Thymic stromal lymphopoietin (TSLP) is known to be elevated and truncated in nasal polyps (NPs) of patients with chronic rhinosinusitis and might play a significant role in type 2 inflammation in this disease. However, neither the structure nor the role of the truncated products of TSLP has been studied. Objective We sought to investigate the mechanisms of truncation of TSLP in NPs and the function of the truncated products. Methods We incubated recombinant human TSLP with NP extracts, and determined the protein sequence of the truncated forms of TSLP using Edman protein sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. We investigated the functional activity of truncated TSLP using a PBMC-based bioassay. Results Edman sequencing and mass spectrometry results indicated that NP extracts generated 2 major truncated products, TSLP (residues 29-124) and TSLP (131-159). Interestingly, these 2 products remained linked with disulfide bonds and presented as a dimerized form, TSLP (29-124 + 131-159). We identified that members of the proprotein convertase were rate-limiting enzymes in the truncation of TSLP between residues 130 and 131 and generated a heterodimeric unstable metabolite TSLP (29-130 + 131-159). Carboxypeptidase N immediately digested 6 amino acids from the C terminus of the longer subunit of TSLP to generate a stable dimerized form, TSLP (29-124 + 131-159), in NPs. These truncations were homeostatic but primate-specific events. A metabolite TSLP (29-130 + 131-159) strongly activated myeloid dendritic cells and group 2 innate lymphoid cells compared with mature TSLP. Conclusions Posttranslational modifications control the functional activity of TSLP in humans and overproduction of TSLP may be a key trigger for the amplification of type 2 inflammation in diseases.",
keywords = "Carboxypeptidase, chronic rhinosinusitis, dendritic cells, group 2 innate lymphoid cells, nasal polyps, posttranslational modification, proprotein convertases, thymic stromal lymphopoietin",
author = "Poposki, {Julie A.} and Klingler, {Aiko I.} and Stevens, {Whitney W.} and Peters, {Anju T.} and Hulse, {Kathryn E.} and Grammer, {Leslie C.} and Schleimer, {Robert P.} and Welch, {Kevin C.} and Smith, {Stephanie S.} and Sidle, {Douglas M.} and Conley, {David B.} and Tan, {Bruce K.} and Kern, {Robert C.} and Atsushi Kato",
year = "2017",
month = "5",
day = "1",
doi = "10.1016/j.jaci.2016.08.040",
language = "English (US)",
volume = "139",
pages = "1559--1567.e8",
journal = "Journal of Allergy and Clinical Immunology",
issn = "0091-6749",
publisher = "Mosby Inc.",
number = "5",

}

TY - JOUR

T1 - Proprotein convertases generate a highly functional heterodimeric form of thymic stromal lymphopoietin in humans

AU - Poposki, Julie A.

AU - Klingler, Aiko I.

AU - Stevens, Whitney W.

AU - Peters, Anju T.

AU - Hulse, Kathryn E.

AU - Grammer, Leslie C.

AU - Schleimer, Robert P.

AU - Welch, Kevin C.

AU - Smith, Stephanie S.

AU - Sidle, Douglas M.

AU - Conley, David B.

AU - Tan, Bruce K.

AU - Kern, Robert C.

AU - Kato, Atsushi

PY - 2017/5/1

Y1 - 2017/5/1

N2 - Rationale Thymic stromal lymphopoietin (TSLP) is known to be elevated and truncated in nasal polyps (NPs) of patients with chronic rhinosinusitis and might play a significant role in type 2 inflammation in this disease. However, neither the structure nor the role of the truncated products of TSLP has been studied. Objective We sought to investigate the mechanisms of truncation of TSLP in NPs and the function of the truncated products. Methods We incubated recombinant human TSLP with NP extracts, and determined the protein sequence of the truncated forms of TSLP using Edman protein sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. We investigated the functional activity of truncated TSLP using a PBMC-based bioassay. Results Edman sequencing and mass spectrometry results indicated that NP extracts generated 2 major truncated products, TSLP (residues 29-124) and TSLP (131-159). Interestingly, these 2 products remained linked with disulfide bonds and presented as a dimerized form, TSLP (29-124 + 131-159). We identified that members of the proprotein convertase were rate-limiting enzymes in the truncation of TSLP between residues 130 and 131 and generated a heterodimeric unstable metabolite TSLP (29-130 + 131-159). Carboxypeptidase N immediately digested 6 amino acids from the C terminus of the longer subunit of TSLP to generate a stable dimerized form, TSLP (29-124 + 131-159), in NPs. These truncations were homeostatic but primate-specific events. A metabolite TSLP (29-130 + 131-159) strongly activated myeloid dendritic cells and group 2 innate lymphoid cells compared with mature TSLP. Conclusions Posttranslational modifications control the functional activity of TSLP in humans and overproduction of TSLP may be a key trigger for the amplification of type 2 inflammation in diseases.

AB - Rationale Thymic stromal lymphopoietin (TSLP) is known to be elevated and truncated in nasal polyps (NPs) of patients with chronic rhinosinusitis and might play a significant role in type 2 inflammation in this disease. However, neither the structure nor the role of the truncated products of TSLP has been studied. Objective We sought to investigate the mechanisms of truncation of TSLP in NPs and the function of the truncated products. Methods We incubated recombinant human TSLP with NP extracts, and determined the protein sequence of the truncated forms of TSLP using Edman protein sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. We investigated the functional activity of truncated TSLP using a PBMC-based bioassay. Results Edman sequencing and mass spectrometry results indicated that NP extracts generated 2 major truncated products, TSLP (residues 29-124) and TSLP (131-159). Interestingly, these 2 products remained linked with disulfide bonds and presented as a dimerized form, TSLP (29-124 + 131-159). We identified that members of the proprotein convertase were rate-limiting enzymes in the truncation of TSLP between residues 130 and 131 and generated a heterodimeric unstable metabolite TSLP (29-130 + 131-159). Carboxypeptidase N immediately digested 6 amino acids from the C terminus of the longer subunit of TSLP to generate a stable dimerized form, TSLP (29-124 + 131-159), in NPs. These truncations were homeostatic but primate-specific events. A metabolite TSLP (29-130 + 131-159) strongly activated myeloid dendritic cells and group 2 innate lymphoid cells compared with mature TSLP. Conclusions Posttranslational modifications control the functional activity of TSLP in humans and overproduction of TSLP may be a key trigger for the amplification of type 2 inflammation in diseases.

KW - Carboxypeptidase

KW - chronic rhinosinusitis

KW - dendritic cells

KW - group 2 innate lymphoid cells

KW - nasal polyps

KW - posttranslational modification

KW - proprotein convertases

KW - thymic stromal lymphopoietin

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U2 - 10.1016/j.jaci.2016.08.040

DO - 10.1016/j.jaci.2016.08.040

M3 - Article

VL - 139

SP - 1559-1567.e8

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

IS - 5

ER -