Discovery of a pyruvylated peptide-metabolizing enzyme using a fluorescent substrate-based protein discovery technique

Kentaro Yoshioka; Toru Komatsu; Kenjiro Hanaoka; Tasuku Ueno; Takuya Terai; Tetsuo Nagano; Yasuteru Urano

doi:10.1039/c6cc00829a

Discovery of a pyruvylated peptide-metabolizing enzyme using a fluorescent substrate-based protein discovery technique

Kentaro Yoshioka, Toru Komatsu, Kenjiro Hanaoka, Tasuku Ueno, Takuya Terai, Tetsuo Nagano, Yasuteru Urano

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

Abstract

We employed a fluorescent substrate-based target discovery approach to screen the enzymome for metabolic activity towards pyruvyl-amidated peptides, and identified an acylamino acid-releasing enzyme (APEH). Cells overexpressing APEH exhibited higher metabolic activity towards the probe, N-pyruvyl-leucyl-7-amido-4-methylcoumarin (Pyr-Leu-AMC), while the selective APEH inhibitor AA74-1 blocked the reaction. Metabolism of various pyruvylated peptides in liver lysate was almost completely blocked by AA74-1. Pyruvyl peptides are synthesized in response to oxidative stress, but their biological role is poorly understood; identification of a key contributor to their metabolism should stimulate research on pathways leading from oxidative stress to protein modification and biological output.

Original language	English
Pages (from-to)	4377-4380
Number of pages	4
Journal	Chemical Communications
Volume	52
Issue number	23
DOIs	https://doi.org/10.1039/c6cc00829a
Publication status	Published - 2016
Externally published	Yes

ASJC Scopus subject areas

Catalysis
Electronic, Optical and Magnetic Materials
Ceramics and Composites
Chemistry(all)
Surfaces, Coatings and Films
Metals and Alloys
Materials Chemistry

Access to Document

10.1039/c6cc00829a

Cite this

@article{babe58913aec4625a4e1904624f38d11,

title = "Discovery of a pyruvylated peptide-metabolizing enzyme using a fluorescent substrate-based protein discovery technique",

abstract = "We employed a fluorescent substrate-based target discovery approach to screen the enzymome for metabolic activity towards pyruvyl-amidated peptides, and identified an acylamino acid-releasing enzyme (APEH). Cells overexpressing APEH exhibited higher metabolic activity towards the probe, N-pyruvyl-leucyl-7-amido-4-methylcoumarin (Pyr-Leu-AMC), while the selective APEH inhibitor AA74-1 blocked the reaction. Metabolism of various pyruvylated peptides in liver lysate was almost completely blocked by AA74-1. Pyruvyl peptides are synthesized in response to oxidative stress, but their biological role is poorly understood; identification of a key contributor to their metabolism should stimulate research on pathways leading from oxidative stress to protein modification and biological output.",

author = "Kentaro Yoshioka and Toru Komatsu and Kenjiro Hanaoka and Tasuku Ueno and Takuya Terai and Tetsuo Nagano and Yasuteru Urano",

note = "Publisher Copyright: {\textcopyright} The Royal Society of Chemistry 2016.",

year = "2016",

doi = "10.1039/c6cc00829a",

language = "English",

volume = "52",

pages = "4377--4380",

journal = "Chemical Communications",

issn = "1359-7345",

publisher = "Royal Society of Chemistry",

number = "23",

}

TY - JOUR

T1 - Discovery of a pyruvylated peptide-metabolizing enzyme using a fluorescent substrate-based protein discovery technique

AU - Yoshioka, Kentaro

AU - Komatsu, Toru

AU - Hanaoka, Kenjiro

AU - Ueno, Tasuku

AU - Terai, Takuya

AU - Nagano, Tetsuo

AU - Urano, Yasuteru

N1 - Publisher Copyright: © The Royal Society of Chemistry 2016.

PY - 2016

Y1 - 2016

N2 - We employed a fluorescent substrate-based target discovery approach to screen the enzymome for metabolic activity towards pyruvyl-amidated peptides, and identified an acylamino acid-releasing enzyme (APEH). Cells overexpressing APEH exhibited higher metabolic activity towards the probe, N-pyruvyl-leucyl-7-amido-4-methylcoumarin (Pyr-Leu-AMC), while the selective APEH inhibitor AA74-1 blocked the reaction. Metabolism of various pyruvylated peptides in liver lysate was almost completely blocked by AA74-1. Pyruvyl peptides are synthesized in response to oxidative stress, but their biological role is poorly understood; identification of a key contributor to their metabolism should stimulate research on pathways leading from oxidative stress to protein modification and biological output.

AB - We employed a fluorescent substrate-based target discovery approach to screen the enzymome for metabolic activity towards pyruvyl-amidated peptides, and identified an acylamino acid-releasing enzyme (APEH). Cells overexpressing APEH exhibited higher metabolic activity towards the probe, N-pyruvyl-leucyl-7-amido-4-methylcoumarin (Pyr-Leu-AMC), while the selective APEH inhibitor AA74-1 blocked the reaction. Metabolism of various pyruvylated peptides in liver lysate was almost completely blocked by AA74-1. Pyruvyl peptides are synthesized in response to oxidative stress, but their biological role is poorly understood; identification of a key contributor to their metabolism should stimulate research on pathways leading from oxidative stress to protein modification and biological output.

UR - http://www.scopus.com/inward/record.url?scp=84960929265&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84960929265&partnerID=8YFLogxK

U2 - 10.1039/c6cc00829a

DO - 10.1039/c6cc00829a

M3 - Article

C2 - 26925595

AN - SCOPUS:84960929265

SN - 1359-7345

VL - 52

SP - 4377

EP - 4380

JO - Chemical Communications

JF - Chemical Communications

IS - 23

ER -

Discovery of a pyruvylated peptide-metabolizing enzyme using a fluorescent substrate-based protein discovery technique

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this