Preliminary structural characterisation of the 33 kDa protein (PsbO) in solution studied by site-directed mutagenesis and NMR spectroscopy

2004 ◽  
Vol 6 (20) ◽  
pp. 4878-4881 ◽  
Author(s):  
Marc Nowaczyk ◽  
Carsten Berghaus ◽  
Raphael Stoll ◽  
Matthias Rögner
Keyword(s):  
Nmr Spectroscopy ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Structural Characterisation

scholarly journals Enzymatic Production of β-Glucose 1,6-Bisphosphate Through Manipulation of Catalytic Magnesium Coordination

2020 ◽  
Author(s):  
Henry P Wood ◽  
Nicola J Baxter ◽  
Clare R Trevitt ◽  
F Aaron Cruz-Navarrete ◽  
Andrea M Hounslow ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Enzymatic Synthesis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Enzymatic Production ◽  
Congenital Disorder Of Glycosylation ◽  
Precipitation Procedure ◽  
Combined Use ◽  
Synthesis Strategy ◽  
Phosphomannomutase 2

<p>Manipulation of enzyme behaviour represents a sustainable technology that can be harnessed to enhance the production of valuable metabolites and chemical precursors. b-glucose 1,6-bisphosphate (bG16BP) is a native reaction intermediate in the catalytic cycle of b-phosphoglucomutase (bPGM) that has been proposed as a treatment for human congenital disorder of glycosylation involving phosphomannomutase 2 (PMM2). Studies of both bPGM and PMM2 could benefit from a green and high-yielding method for bG16BP production. Three strategies have been reported previously for the synthesis of bG16BP; however, each of these methods either delivers low yields or uses chemicals and procedures with significant environmental impacts. Herein, through combined use of NMR spectroscopy, kinetic assays and site-directed mutagenesis, we report the efficient enzymatic synthesis of anomer-specific bG16BP using a variant of bPGM. Further purification, employing a simple environmentally considerate precipitation procedure requiring only a standard biochemical toolset, results in a product with high purity and yield. Moreover, this synthesis strategy illustrates how manipulation of the catalytic magnesium coordination of an enzyme can be utilised to generate large quantities of a valuable metabolite.</p>

Site-directed mutagenesis and proton NMR spectroscopy of an interdomain segment in the pyruvate dehydrogenase multienzyme complex of Escherichia coli

Biochemistry ◽  
1988 ◽  
Vol 27 (1) ◽  
pp. 289-296 ◽  
Author(s):  
Frieda L. Texter ◽  
Sheena E. Radford ◽  
Ernest D. Laue ◽  
Richard N. Perham ◽  
John S. Miles ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nmr Spectroscopy ◽  
Pyruvate Dehydrogenase ◽  
Proton Nmr ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Multienzyme Complex ◽  
Proton Nmr Spectroscopy

scholarly journals Determination of pKavalues of the histidine side chains of phosphatidylinositol-specific phospholipase C fromBacillus cereusby NMR spectroscopy and site-directed mutagenesis

1997 ◽  
Vol 6 (9) ◽  
pp. 1937-1944 ◽  
Author(s):  
Tun Liu ◽  
Margret Ryan ◽  
Frederick W. Dahlquist ◽  
O. Hayes Griffith
Keyword(s):  
Nmr Spectroscopy ◽  
Phospholipase C ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Side Chains

Site-directed mutagenesis and high-resolution NMR spectroscopy of the active site of porphobilinogen deaminase

Biochemistry ◽  
1988 ◽  
Vol 27 (21) ◽  
pp. 7984-7990 ◽  
Author(s):  
A. I. Scott ◽  
C. A. Roessner ◽  
N. J. Stolowich ◽  
P. Karuso ◽  
H. J. Williams ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
High Resolution ◽  
Active Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Porphobilinogen Deaminase ◽  
High Resolution Nmr

scholarly journals Aspects of Receptor Binding and Signalling of Interleukin-4 Investigated by Site-directed Mutagenesis and NMR Spectroscopy

1994 ◽  
Vol 237 (4) ◽  
pp. 423-436 ◽  
Author(s):  
Thomas Müller ◽  
Thorsten Dieckmann ◽  
Walter Sebald ◽  
Hartmut Oschkinat
Keyword(s):  
Nmr Spectroscopy ◽  
Receptor Binding ◽  
Site Directed Mutagenesis ◽  
Interleukin 4 ◽  
Directed Mutagenesis

scholarly journals Enzymatic Production of β-Glucose 1,6-Bisphosphate Through Manipulation of Catalytic Magnesium Coordination

2020 ◽  
Author(s):  
Henry P Wood ◽  
Nicola J Baxter ◽  
Clare R Trevitt ◽  
F Aaron Cruz-Navarrete ◽  
Andrea M Hounslow ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Enzymatic Synthesis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Enzymatic Production ◽  
Congenital Disorder Of Glycosylation ◽  
Precipitation Procedure ◽  
Combined Use ◽  
Synthesis Strategy ◽  
Phosphomannomutase 2

<p>Manipulation of enzyme behaviour represents a sustainable technology that can be harnessed to enhance the production of valuable metabolites and chemical precursors. b-glucose 1,6-bisphosphate (bG16BP) is a native reaction intermediate in the catalytic cycle of b-phosphoglucomutase (bPGM) that has been proposed as a treatment for human congenital disorder of glycosylation involving phosphomannomutase 2 (PMM2). Studies of both bPGM and PMM2 could benefit from a green and high-yielding method for bG16BP production. Three strategies have been reported previously for the synthesis of bG16BP; however, each of these methods either delivers low yields or uses chemicals and procedures with significant environmental impacts. Herein, through combined use of NMR spectroscopy, kinetic assays and site-directed mutagenesis, we report the efficient enzymatic synthesis of anomer-specific bG16BP using a variant of bPGM. Further purification, employing a simple environmentally considerate precipitation procedure requiring only a standard biochemical toolset, results in a product with high purity and yield. Moreover, this synthesis strategy illustrates how manipulation of the catalytic magnesium coordination of an enzyme can be utilised to generate large quantities of a valuable metabolite.</p>

Location of the Zn2+-Binding Site on S100B As Determined by NMR Spectroscopy and Site-Directed Mutagenesis†

Biochemistry ◽  
2003 ◽  
Vol 42 (46) ◽  
pp. 13410-13421 ◽  
Author(s):  
Paul T. Wilder ◽  
Donna M. Baldisseri ◽  
Ryan Udan ◽  
Kristen M. Vallely ◽  
David J. Weber
Keyword(s):  
Nmr Spectroscopy ◽  
Binding Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis
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