Substrate-induced closure of the flap domain in the ternary complex structures provides insights into the mechanism of catalysis by 3-hydroxy-3-methylglutaryl–CoA reductase

Tabernero, Lydia; Bochar, Daniel A.; Rodwell, Victor W.; Stauffacher, Cynthia V.

doi:10.1073/pnas.96.13.7167

Cited by 87 publications

(187 citation statements)

References 18 publications

Supporting

Mentioning

176

Contrasting

Order By: Relevance

“…Dimerization of the catalytic domains is critical for HMGCoA reductase activity, because the enzyme active site is formed by interactions between two monomers (Lawrence et al, 1995;Rogers et al, 1997;Tabernero et al, 1999). Recent results also support the importance of oligomerization for regulated degradation of HMG-CoA reductase, such that monomeric fusion proteins are degraded rapidly relative to fusion proteins capable of oligomerizing via their carboxyl termini (Cheng et al, 1999).…”

Section: Discussionmentioning

confidence: 66%

“…In eukaryotes, the carboxyl terminus of HMG-CoA reductase contains the cytosolic catalytic domain, and the amino terminus contains the membrane domain (Liscum et al, 1985). Although details concerning the structure and function of the HMG-CoA reductase catalytic domain are beginning to be solved (Darnay et al, 1992;Lawrence et al, 1995;Rogers et al, 1997;Tabernero et al, 1999), the HMG-CoA reductase membrane domain remains much more cryptic. In plants, the membrane domain contains two transmembrane helices (Denbow et al, 1996;Re et al, 1997), whereas in animals and fungi, it contains of a series of seven or eight transmembrane helices (Liscum et al, 1985;Basson et al, 1988;Sengstag et al, 1990;Olender and Simon, 1992;Roitelman et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

“…To test this idea, we examined karmellae assembly in a strain expressing both full-length Hmg1p and a truncated protein containing only the linker and the catalytic domain of Hmg1p. Because HMG-CoA reductase is thought to act as a dimer using contact sites in the catalytic domain (Edwards et al, 1985;Basson et al, 1987;Frimpong and Rodwell, 1994;Lawrence et al, 1995;Rogers et al, 1997;Tabernero et al, 1999), we hypothesized that expression of a soluble catalytic domain would interfere with the ability of the holoprotein to dimerize and to induce karmellae. It is important to note that the catalytic domain is capable of proper folding and oligomerization in the absence of attachment to the membrane domain, because it is enzymatically active,and the amount of activity parallels the protein amount (Donald et al, 1997).…”

Section: Competition With the Soluble Catalytic Domain Reduced The Ovmentioning

confidence: 99%

See 2 more Smart Citations

The Role of the 3-Hydroxy 3-Methylglutaryl Coenzyme A Reductase Cytosolic Domain in Karmellae Biogenesis

Profant¹,

Roberts

Koning

et al. 1999

MBoC

View full text Add to dashboard Cite

In all cells examined, specific endoplasmic reticulum (ER) membrane arrays are induced in response to increased levels of the ER membrane protein 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase. In yeast, expression of Hmg1p, one of two yeast HMG-CoA reductase isozymes, induces assembly of nuclear-associated ER stacks called karmellae. Understanding the features of HMG-CoA reductase that signal karmellae biogenesis would provide useful insights into the regulation of membrane biogenesis. The HMG-CoA reductase protein consists of two domains, a multitopic membrane domain and a cytosolic catalytic domain. Previous studies had indicated that the HMG-CoA reductase membrane domain was exclusively responsible for generation of ER membrane proliferations. Surprisingly, we discovered that this conclusion was incorrect: sequences at the carboxyl terminus of HMG-CoA reductase can profoundly affect karmellae biogenesis. Specifically, truncations of Hmg1p that removed or shortened the carboxyl terminus were unable to induce karmellae assembly. This result indicated that the membrane domain of Hmg1p was not sufficient to signal for karmellae assembly. Using β-galactosidase fusions, we demonstrated that the carboxyl terminus was unlikely to simply serve as an oligomerization domain. Our working hypothesis is that a truncated or misfolded cytosolic domain prevents proper signaling for karmellae by interfering with the required tertiary structure of the membrane domain.

show abstract

Section: Discussionmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Competition With the Soluble Catalytic Domain Reduced The Ovmentioning

confidence: 99%

See 1 more Smart Citation

The Role of the 3-Hydroxy 3-Methylglutaryl Coenzyme A Reductase Cytosolic Domain in Karmellae Biogenesis

Profant¹,

Roberts

Koning

et al. 1999

MBoC

View full text Add to dashboard Cite

show abstract

“…Although prior investigations have established the probable functions of an active site glutamate, aspartate, histidine, and lysine that participate in a common mechanism of catalysis~Wang et al ., 1990;Darnay et al, 1992;Darnay & Rodwell, 1993;Frimpong & Rodwell, 1994;Tabernero et al, 1998;Bochar et al, 1999c!, only recently was it possible to infer from gene sequences that there are two structural classes of HMG-CoA reductase~Doo-little & Logsdon, 1998; Bochar et al, 1999b!. Almost all prior research has addressed the class I enzymes of higher vertebrates that constitute the target for the inhibitory statin drugs employed to control cholesterol biosynthesis in human subjects.…”

Section: ! 4!mentioning

confidence: 99%

Dual coenzyme specificity of Archaeoglobus fulgidus HMG‐CoA reductase

2000

Self Cite

View full text Add to dashboard Cite

Comparison of the inferred amino acid sequence of orf AF1736 of Archaeoglobus fulgidus to that of Pseudomonas mevalonii HMG-CoA reductase suggested that AF1736 might encode a Class II HMG-CoA reductase. Following polymerase chain reaction-based cloning of AF1736 from A. fulgidus genomic DNA and expression in Escherichia coli, the encoded enzyme was purified to apparent homogeneity and its enzymic properties were determined. Activity was optimal at 85 8C, DH a was 54 kJ0mol, and the statin drug mevinolin inhibited competitively with HMG-CoÃ K i 180 mM!. Protonated forms of His390 and Lys277, the apparent cognates of the active site histidine and lysine of the P. mevalonii enzyme, appear essential for activity. The mechanism proposed for catalysis of P. mevalonii HMG-CoA reductase thus appears valid for A. fulgidus HMG-CoA reductase. Unlike any other HMG-CoA reductase, the A. fulgidus enzyme exhibits dual coenzyme specificity. pH-activity profiles for all four reactions revealed that optimal activity using NADP~H! occurred at a pH from 1 to 3 units more acidic than that observed using NAD~H!. Kinetic parameters were therefore determined for all substrates for all four catalyzed reactions using either NAD~H! or NADP~H!. NADPH and NADH compete for occupancy of a common site. k cat @NAD~H!#0k cat @NADP~H!# varied from unity to under 70 for the four reactions, indicative of slight preference for NAD~H!. The results indicate the importance of the protonated status of active site residues His390 and Lys277, shown by altered K M and k cat values, and indicate that NAD~H! and NADP~H! have comparable affinity for the same site.Keywords: archaeal oxidoreductase; HMG-CoA reductase; mevaldehyde; mevalonate; redox coenzyme specificity; thermostable enzyme Catalysis by HMG-CoA reductase~E.C. 1.1.1.34! of Reaction~1!, the four-electron reductive deacylation of~S!-HMG-CoA to the isoprenoid precursor~R!-mevalonate, proceeds in three stages, the first and third of which are reductive. The putative intermediates mevaldyl-CoA and mevaldehyde remain enzyme-bound during the course of the reaction. HMG-CoA reductase also catalyzes Reactions 2! and~3!, two 2-electron reactions of free mevaldehyde that appear to model the third stage and the reverse of the second and first stages of Reaction~1!, respectively, and Reaction~4!, the four-electron oxidative acylation of~R!-mevalonate to~S!-HMG-CoA, the reverse of Reaction~1!~Bochar et al., 1999a!.~1 ! Reprint requests to: Victor W. Rodwell, Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907; e-mail: vrodwell@purdue.edu. Abbreviations: CoASH, coenzyme A; HMG, 3-hydroxy-3-methylglutaryl; PCR, polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The suffixes P and A refer to residues from the HMG-CoA reductases of Pseudomonas mevalonii and of Archaeoglobus fulgidus, respectively.

show abstract

“…The first example considers a subunit of the homodimer 3-hydroxy-3-methylglutaryl-CoA reductase (Figure 1), an enzyme involved in the biosynthesis of cholesterol in mammals (Tabernero et al, 1999). Two clusters are predicted for this example, one showing a coverage of 38.13% and precision of 79.1% and the other showing a coverage of 25.9% and a precision of 66.67%.…”

Section: Resultsmentioning

confidence: 99%

A simple and efficient method for predicting protein-protein interaction sites

Higa

Tozzi

2008

Genet. Mol. Res.

View full text Add to dashboard Cite

AbstrAct.Computational methods for predicting protein-protein interaction sites based on structural data are characterized by an accuracy between 70 and 80%. Some experimental studies indicate that only a fraction of the residues, forming clusters in the center of the interaction site, are energetically important for binding. In addition, the analysis of amino acid composition has shown that residues located in the center of the interaction site can be better discriminated from the residues in other parts of the protein surface. In the present study, we implement a simple method to predict interaction site residues exploiting this fact and show that it achieves a very competitive performance compared to other methods using the same dataset and criteria for performance evaluation (success rate of 82.1%).

show abstract

Substrate-induced closure of the flap domain in the ternary complex structures provides insights into the mechanism of catalysis by 3-hydroxy-3-methylglutaryl–CoA reductase

Cited by 87 publications

References 18 publications

The Role of the 3-Hydroxy 3-Methylglutaryl Coenzyme A Reductase Cytosolic Domain in Karmellae Biogenesis

The Role of the 3-Hydroxy 3-Methylglutaryl Coenzyme A Reductase Cytosolic Domain in Karmellae Biogenesis

Dual coenzyme specificity of Archaeoglobus fulgidus HMG‐CoA reductase

A simple and efficient method for predicting protein-protein interaction sites

Contact Info

Product

Resources

About