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.
| Original language | English |
|---|---|
| Pages (from-to) | 203-206 |
| Number of pages | 4 |
| Journal | FEBS Letters |
| Volume | 528 |
| Issue number | 1-3 |
| DOIs | |
| Publication status | Published - 2002 Sep 25 |
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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
Cite this
NMR studies of the hydrogen bonds involving the catalytic triad of Escherichia coli thioesterase/protease I. / Tyukhtenko, Sergiy I.; Litvinchuk, Alexandra V.; Chang, Chi Fon; Leu, Ruey Jyh; Shaw, Jei Fu; Huang, Tai-huang.
In: FEBS Letters, Vol. 528, No. 1-3, 25.09.2002, p. 203-206.Research output: Contribution to journal › Article
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TY - JOUR
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
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
UR - http://www.scopus.com/inward/record.url?scp=0037174164&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037174164&partnerID=8YFLogxK
U2 - 10.1016/S0014-5793(02)03308-2
DO - 10.1016/S0014-5793(02)03308-2
M3 - Article
C2 - 12297305
AN - SCOPUS:0037174164
VL - 528
SP - 203
EP - 206
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 1-3
ER -