The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads

Ki Hyun Nam; Soo Jin Kim; Amit Priyadarshi; Hyun Sook Kim; Kwang Yeon Hwang

doi:10.1016/j.bbrc.2009.08.123

The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads

Ki Hyun Nam
, Soo Jin Kim
, Amit Priyadarshi
, Hyun Sook Kim
, Kwang Yeon Hwang^*

^*Corresponding author for this work

Department of Biosystems and Biotechnology

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

26 Citations (Scopus)

Abstract

The esterase/lipase family (EC 3.1.1.3/EC 3.1.1.1) represents a diverse group of hydrolases that catalyze the cleavage of ester bonds and are widely distributed in animals, plants and microorganisms. Among these enzymes, hormone-sensitive lipases, play a critical role in the regulation of rodent fat cell lipolysis and are regarded as adipose tissue-specific enzymes. Recently, we reported the structural and biological characterization of EstE5 from the metagenome library [K.H. Nam, M.Y. Kim, S.J. Kim, A. Priyadarshi, W.H. Lee, K.Y. Hwang, Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase, Biochem. Biophys. Res. Commun. 379 (2009) 553-556]. The structure of this protein revealed that it belongs to the HSL-family. Here, we report the inhibition of the activity of the HSL-homolog EstE5 protein as determined by the use of esterase/lipase inhibitors. Our results revealed that the EstE5 protein is significantly inhibited by PMSF. In addition, this is the first study to identify the crystal structures of EstE5-PMSF at 2.4 and 2.5 Å among the HSL-homolog structures. This structural configuration is similar to that adopted when serine proteases are inhibited by PMSF. The results presented here provide valuable information regarding the properties of the HSL-family.

Original language	English
Pages (from-to)	247-250
Number of pages	4
Journal	Biochemical and biophysical research communications
Volume	389
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2009.08.123
Publication status	Published - 2009 Nov 13

Bibliographical note

Funding Information:
We thank H.S. Lee and his staff for assistance during the data collection at beamline 4A of the Pohang Light Source, Korea. This study was supported by the Functional Proteomics Center, 21C Frontier Program, of the Korea Ministry of Science and Technology ( M108KM010031-08K1301-03111 ) and the Korea Science and Engineering Foundation ; Grant No. ( R01-2007-000-20072-0 (2008)). K.H. Nam is supported by the Brain Korea 21 project.

Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.

Keywords

Active site inhibition
Charge-relay system
EstE5
EstE5-PMSF
Esterase/lipase

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2009.08.123

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The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads. / Nam, Ki Hyun; Kim, Soo Jin; Priyadarshi, Amit et al.
In: Biochemical and biophysical research communications, Vol. 389, No. 2, 13.11.2009, p. 247-250.

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

@article{4b8301a95144451ba2b02c2e302f8bd4,

title = "The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads",

abstract = "The esterase/lipase family (EC 3.1.1.3/EC 3.1.1.1) represents a diverse group of hydrolases that catalyze the cleavage of ester bonds and are widely distributed in animals, plants and microorganisms. Among these enzymes, hormone-sensitive lipases, play a critical role in the regulation of rodent fat cell lipolysis and are regarded as adipose tissue-specific enzymes. Recently, we reported the structural and biological characterization of EstE5 from the metagenome library [K.H. Nam, M.Y. Kim, S.J. Kim, A. Priyadarshi, W.H. Lee, K.Y. Hwang, Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase, Biochem. Biophys. Res. Commun. 379 (2009) 553-556]. The structure of this protein revealed that it belongs to the HSL-family. Here, we report the inhibition of the activity of the HSL-homolog EstE5 protein as determined by the use of esterase/lipase inhibitors. Our results revealed that the EstE5 protein is significantly inhibited by PMSF. In addition, this is the first study to identify the crystal structures of EstE5-PMSF at 2.4 and 2.5 {\AA} among the HSL-homolog structures. This structural configuration is similar to that adopted when serine proteases are inhibited by PMSF. The results presented here provide valuable information regarding the properties of the HSL-family.",

keywords = "Active site inhibition, Charge-relay system, EstE5, EstE5-PMSF, Esterase/lipase",

author = "Nam, \{Ki Hyun\} and Kim, \{Soo Jin\} and Amit Priyadarshi and Kim, \{Hyun Sook\} and Hwang, \{Kwang Yeon\}",

note = "Funding Information: We thank H.S. Lee and his staff for assistance during the data collection at beamline 4A of the Pohang Light Source, Korea. This study was supported by the Functional Proteomics Center, 21C Frontier Program, of the Korea Ministry of Science and Technology ( M108KM010031-08K1301-03111 ) and the Korea Science and Engineering Foundation ; Grant No. ( R01-2007-000-20072-0 (2008)). K.H. Nam is supported by the Brain Korea 21 project. Copyright: Copyright 2011 Elsevier B.V., All rights reserved.",

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doi = "10.1016/j.bbrc.2009.08.123",

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AU - Kim, Soo Jin

AU - Priyadarshi, Amit

AU - Kim, Hyun Sook

AU - Hwang, Kwang Yeon

N1 - Funding Information: We thank H.S. Lee and his staff for assistance during the data collection at beamline 4A of the Pohang Light Source, Korea. This study was supported by the Functional Proteomics Center, 21C Frontier Program, of the Korea Ministry of Science and Technology ( M108KM010031-08K1301-03111 ) and the Korea Science and Engineering Foundation ; Grant No. ( R01-2007-000-20072-0 (2008)). K.H. Nam is supported by the Brain Korea 21 project. Copyright: Copyright 2011 Elsevier B.V., All rights reserved.

PY - 2009/11/13

Y1 - 2009/11/13

N2 - The esterase/lipase family (EC 3.1.1.3/EC 3.1.1.1) represents a diverse group of hydrolases that catalyze the cleavage of ester bonds and are widely distributed in animals, plants and microorganisms. Among these enzymes, hormone-sensitive lipases, play a critical role in the regulation of rodent fat cell lipolysis and are regarded as adipose tissue-specific enzymes. Recently, we reported the structural and biological characterization of EstE5 from the metagenome library [K.H. Nam, M.Y. Kim, S.J. Kim, A. Priyadarshi, W.H. Lee, K.Y. Hwang, Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase, Biochem. Biophys. Res. Commun. 379 (2009) 553-556]. The structure of this protein revealed that it belongs to the HSL-family. Here, we report the inhibition of the activity of the HSL-homolog EstE5 protein as determined by the use of esterase/lipase inhibitors. Our results revealed that the EstE5 protein is significantly inhibited by PMSF. In addition, this is the first study to identify the crystal structures of EstE5-PMSF at 2.4 and 2.5 Å among the HSL-homolog structures. This structural configuration is similar to that adopted when serine proteases are inhibited by PMSF. The results presented here provide valuable information regarding the properties of the HSL-family.

AB - The esterase/lipase family (EC 3.1.1.3/EC 3.1.1.1) represents a diverse group of hydrolases that catalyze the cleavage of ester bonds and are widely distributed in animals, plants and microorganisms. Among these enzymes, hormone-sensitive lipases, play a critical role in the regulation of rodent fat cell lipolysis and are regarded as adipose tissue-specific enzymes. Recently, we reported the structural and biological characterization of EstE5 from the metagenome library [K.H. Nam, M.Y. Kim, S.J. Kim, A. Priyadarshi, W.H. Lee, K.Y. Hwang, Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase, Biochem. Biophys. Res. Commun. 379 (2009) 553-556]. The structure of this protein revealed that it belongs to the HSL-family. Here, we report the inhibition of the activity of the HSL-homolog EstE5 protein as determined by the use of esterase/lipase inhibitors. Our results revealed that the EstE5 protein is significantly inhibited by PMSF. In addition, this is the first study to identify the crystal structures of EstE5-PMSF at 2.4 and 2.5 Å among the HSL-homolog structures. This structural configuration is similar to that adopted when serine proteases are inhibited by PMSF. The results presented here provide valuable information regarding the properties of the HSL-family.

KW - Active site inhibition

KW - Charge-relay system

KW - EstE5

KW - EstE5-PMSF

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SN - 0006-291X

VL - 389

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JO - Biochemical and biophysical research communications

JF - Biochemical and biophysical research communications

IS - 2

ER -

The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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