Crystal structure of Pseudomonas mevalonii HMG-CoA reductase at 3.0 angstrom resolution

Science ◽  
1995 ◽  
Vol 268 (5218) ◽  
pp. 1758-1762 ◽  
Author(s):  
C. Lawrence ◽  
V. Rodwell ◽  
C. Stauffacher
Keyword(s):  
Crystal Structure ◽  
Hmg Coa Reductase ◽  
Pseudomonas Mevalonii ◽  
Coa Reductase

scholarly journals pH dependent inhibition from ammonium ions in the Pseudomonas mevalonii HMG-CoA Reductase crystallization environment

2020 ◽  
Author(s):  
Vatsal Purohit ◽  
C. Nicklaus Steussy ◽  
Anthony R. Rosales ◽  
Chandra J. Critchelow ◽  
Tim Schmidt ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Pathogenic Bacteria ◽  
Coenzyme A ◽  
Enzymatic Reaction ◽  
Hydride Transfer ◽  
Reverse Direction ◽  
Ammonium Ions ◽  
Hmg Coa Reductase ◽  
Pseudomonas Mevalonii ◽  
Coa Reductase

ABSTRACTHMG-CoA reductase (Pseudomonas mevalonii) utilizes mevalonate, coenzyme A (CoA) and the cofactor NAD in a complex mechanism involving two hydride transfers with cofactor exchange, accompanied by large conformational changes by a 50 residue subdomain, to generate HMG-CoA. Details about this mechanism such as the conformational changes that allow intermediate formation, cofactor exchange and product release remain unknown. The formation of the proposed intermediates has also not been observed in structural studies with natural substrates. Having been shown to be an essential enzyme for the survival of gram-positive antibiotic resistant pathogenic bacteria, studying its mechanism in detail will be beneficial in developing novel antibacterials. The enzyme has been shown to be catalytically active inside the crystal with dithio-HMG-CoA and NADH but curiously is found to be inactive in the reverse direction in the structure bound to mevalonate, CoA and NAD.To understand the factors limiting activity in the HMGR crystal with mevalonate, CoA and NAD, we studied the effect of crystallization components and pH on enzymatic activity. We observed a strong inhibition in the crystallization buffer and an increase in activity with increasing pH. We attribute this inhibitive effect to the presence of ammonium ions present in the crystal since inhibition is also observed with several other ammonium salt buffers. Additionally, the lack of inhibition was observed in the absence of ammonium. The effect of each ligand (mevalonate, CoA and NAD) on the rate of the enzymatic reaction in the crystallization environment was further investigated by measuring their Km in the crystallization buffer. The Km measurements indicate that the hydride transfer step between NAD and mevalonate is inhibited in the crystallization environment. To test this further, we solved a crystal structure of pmHMGR bound to the post-hydride transfer intermediate (mevaldehyde) and cofactor Coenzyme A. The resulting turnover with the formation of a thiohemiacetal indicated that the crystallization environment inhibited the oxidative acylation of mevalonate and the reaction intermediate mevaldyl-CoA.

scholarly journals The crystal structure of HMG-CoA reductase fromPseudomonas mevalonii

1993 ◽  
Vol 49 (s1) ◽  
pp. c90-c91
Author(s):  
C. M. Lawrence ◽  
V. W. Rodwell ◽  
C. V. Stauffacher
Keyword(s):  
Crystal Structure ◽  
Hmg Coa Reductase ◽  
Coa Reductase

scholarly journals Crystal structure of the catalytic portion of human HMG-CoA reductase: insights into regulation of activity and catalysis

2000 ◽  
Vol 19 (5) ◽  
pp. 819-830 ◽  
Author(s):  
Eva S. Istvan ◽  
Maya Palnitkar ◽  
Susan K. Buchanan ◽  
Johann Deisenhofer
Keyword(s):  
Crystal Structure ◽  
Hmg Coa Reductase ◽  
Regulation Of Activity ◽  
Coa Reductase

Reduction of HMG-CoA-reductase leads to improved cardiac function in doxorubicin-induced cardiomyopathy in mice

2008 ◽  
Vol 7 ◽  
pp. 202-203
Author(s):  
A RIAD ◽  
S BIEN ◽  
F ESCHER ◽  
D WESTERMANN ◽  
U LANDMESSER ◽  
...  
Keyword(s):  
Cardiac Function ◽  
Hmg Coa Reductase ◽  
Coa Reductase
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