NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I

Sergiy I. Tyukhtenko; Alexandra V. Litvinchuk; Chi Fon Chang; Ruey Jyh Leu; Jei Fu Shaw; Tai Huang Huang

doi:10.1016/S0014-5793(02)03308-2

NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I

Sergiy I. Tyukhtenko, Alexandra V. Litvinchuk, Chi Fon Chang, Ruey Jyh Leu, Jei Fu Shaw, Tai Huang Huang

Department of Physics

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

14 Citations (Scopus)

Abstract

Escherichia coli thioesterase/protease I (TEP-I) is a lipolytic enzyme of the serine protease superfamily with Ser¹⁰, Asp¹⁵⁴ and His¹⁵⁷ as the catalytic triad residues. Based on comparison of the low-field ¹H nuclear magnetic resonance spectra of two mutants (S10G and S12G) and two transition state analogue complexes we have assigned the exchangeable proton resonances at 16.3 ppm, 14.3 ppm, and 12.8 ppm at pH 3.5 to His¹⁵⁷-N^δ1H, Ser¹⁰-O^γH and His¹⁵⁷-N^ε2H, respectively. Thus, the presence of a strong Asp¹⁵⁴-His¹⁵⁷ hydrogen bond in free TEP-I was observed. However, Ser¹⁰-O^γH was shown to form a H-bond with a residue other than His¹⁵⁷-N^ε2.

Original language	English
Pages (from-to)	203-206
Number of pages	4
Journal	FEBS Letters
Volume	528
Issue number	1-3
DOIs	https://doi.org/10.1016/S0014-5793(02)03308-2
Publication status	Published - 2002 Sep 25

Keywords

Lipolytic enzyme
Low barrier hydrogen bond
Nuclear magnetic resonance
Serine protease
Thioesterase

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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title = "NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I",

abstract = "Escherichia coli thioesterase/protease I (TEP-I) is a lipolytic enzyme of the serine protease superfamily with Ser10, Asp154 and His157 as the catalytic triad residues. Based on comparison of the low-field 1H nuclear magnetic resonance spectra of two mutants (S10G and S12G) and two transition state analogue complexes we have assigned the exchangeable proton resonances at 16.3 ppm, 14.3 ppm, and 12.8 ppm at pH 3.5 to His157-Nδ1H, Ser10-OγH and His157-Nε2H, respectively. Thus, the presence of a strong Asp154-His157 hydrogen bond in free TEP-I was observed. However, Ser10-OγH was shown to form a H-bond with a residue other than His157-Nε2.",

keywords = "Lipolytic enzyme, Low barrier hydrogen bond, Nuclear magnetic resonance, Serine protease, Thioesterase",

author = "Tyukhtenko, {Sergiy I.} and Litvinchuk, {Alexandra V.} and Chang, {Chi Fon} and Leu, {Ruey Jyh} and Shaw, {Jei Fu} and Huang, {Tai Huang}",

note = "Funding Information: We thank Dr. Yen-Chyan Liaw for the use of the X-ray crystal structure coordinates of TEP-I prior to publication. This work was supported by Grants from the National Science Council (NSC91-2113-M-001-038) (T.-H.H.) and Academia Sinica (T.-H.H. and J.-F.S.) of the Republic of China. ",

year = "2002",

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doi = "10.1016/S0014-5793(02)03308-2",

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pages = "203--206",

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T1 - NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I

AU - Tyukhtenko, Sergiy I.

AU - Litvinchuk, Alexandra V.

AU - Chang, Chi Fon

AU - Leu, Ruey Jyh

AU - Shaw, Jei Fu

AU - Huang, Tai Huang

N1 - Funding Information: We thank Dr. Yen-Chyan Liaw for the use of the X-ray crystal structure coordinates of TEP-I prior to publication. This work was supported by Grants from the National Science Council (NSC91-2113-M-001-038) (T.-H.H.) and Academia Sinica (T.-H.H. and J.-F.S.) of the Republic of China.

PY - 2002/9/25

Y1 - 2002/9/25

N2 - Escherichia coli thioesterase/protease I (TEP-I) is a lipolytic enzyme of the serine protease superfamily with Ser10, Asp154 and His157 as the catalytic triad residues. Based on comparison of the low-field 1H nuclear magnetic resonance spectra of two mutants (S10G and S12G) and two transition state analogue complexes we have assigned the exchangeable proton resonances at 16.3 ppm, 14.3 ppm, and 12.8 ppm at pH 3.5 to His157-Nδ1H, Ser10-OγH and His157-Nε2H, respectively. Thus, the presence of a strong Asp154-His157 hydrogen bond in free TEP-I was observed. However, Ser10-OγH was shown to form a H-bond with a residue other than His157-Nε2.

AB - Escherichia coli thioesterase/protease I (TEP-I) is a lipolytic enzyme of the serine protease superfamily with Ser10, Asp154 and His157 as the catalytic triad residues. Based on comparison of the low-field 1H nuclear magnetic resonance spectra of two mutants (S10G and S12G) and two transition state analogue complexes we have assigned the exchangeable proton resonances at 16.3 ppm, 14.3 ppm, and 12.8 ppm at pH 3.5 to His157-Nδ1H, Ser10-OγH and His157-Nε2H, respectively. Thus, the presence of a strong Asp154-His157 hydrogen bond in free TEP-I was observed. However, Ser10-OγH was shown to form a H-bond with a residue other than His157-Nε2.

KW - Lipolytic enzyme

KW - Low barrier hydrogen bond

KW - Nuclear magnetic resonance

KW - Serine protease

KW - Thioesterase

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U2 - 10.1016/S0014-5793(02)03308-2

DO - 10.1016/S0014-5793(02)03308-2

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AN - SCOPUS:0037174164

VL - 528

SP - 203

EP - 206

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-3

ER -

NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I

Abstract

Keywords

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