On the Mechanism of Formation of Isopentenylpyrophosphate<sup>*</sup>

Lindberg, Marjorie C.; Yuan, Cong; deWaard, A.; Bloch, Konrad

doi:10.1021/bi00907a027

Cited by 38 publications

(12 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Arg144 positions the substrate in the active site [Fig. (C)], and the Asp283/Lys17 pair has been proposed to directly deprotonate the 3‐OH of MVAPP, triggering phosphorylation . The catalytic role for the invariant Asp/Lys pair of MDD is supported by its active site position and previous mechanistic studies, which showed that a D283A mutant yields a 10 5 fold reduction in activity, and a K17A mutant yields no detectable activity …”

Section: Resultsmentioning

confidence: 95%

“…The phosphorylation stage involves transfer of the γ‐phosphate of ATP to the 3‐OH position of MVAPP, generating the intermediate, 3‐phospho‐mevalonate 5‐diphosphate (3P‐MVAPP) [Fig. (A)] . This intermediate was originally thought to spontaneously decompose to IPP, CO 2 and PO 4 due to the instability of the sterically hindered tertiary phosphate, but we recently reported the analogous tertiary phosphorylated intermediates, mevalonate 3‐phosphate and mevalonate 3,5‐bisphosphate are both stable .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

et al. 2014

View full text Add to dashboard Cite

In animals, cholesterol is made from 5-carbon building blocks produced by the mevalonate pathway. Drugs that inhibit the mevalonate pathway such as atorvastatin (lipitor) have led to successful treatments for high cholesterol in humans. Another potential target for the inhibition of cholesterol synthesis is mevalonate diphosphate decarboxylase (MDD), which catalyzes the phosphorylation of (R)-mevalonate diphosphate, followed by decarboxylation to yield isopentenyl pyrophosphate. We recently discovered an MDD homolog, mevalonate-3-kinase (M3K) from Thermoplasma acidophilum, which catalyzes the identical phosphorylation of (R)-mevalonate, but without concomitant decarboxylation. Thus, M3K catalyzes half the reaction of the decarboxylase, allowing us to separate features of the active site that are required for decarboxylation from features required for phosphorylation. Here we determine the crystal structure of M3K in the apo form, and with bound substrates, and compare it to MDD structures. Structural and mutagenic analysis reveals modifications that allow M3K to bind mevalonate rather than mevalonate diphosphate. Comparison to homologous MDD structures show that both enzymes employ analogous Arg or Lys residues to catalyze phosphate transfer. However, an invariant active site Asp/Lys pair of MDD previously thought to play a role in phosphorylation is missing in M3K with no functional replacement. Thus, we suggest that the invariant Asp/Lys pair in MDD may be critical for decarboxylation rather than phosphorylation.

show abstract

Section: Resultsmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

et al. 2014

View full text Add to dashboard Cite

show abstract

“…30,31 In a previous mechanistic study of mevalonate pyrophosphate decarboxylase, a heavy oxygen atom was incorporated into mevalonate pyrophosphate at the 3-OH position, and then after ATP-dependent conversion to IPP, the heavy oxygen was detected in free phosphate. 30 This suggests a simple mechanism in which the γ-phosphate from ATP is transferred to the 3-OH position of mevalonate pyrophosphate, activating the substrate for decarboxylation. The current literature predicts this tertiary phosphorylated molecule is inherently unstable and falls apart with concomitant decarboxylation.…”

Section: Discussionmentioning

confidence: 99%

Evidence of a Novel Mevalonate Pathway in Archaea

et al. 2014

View full text Add to dashboard Cite

Isoprenoids make up a remarkably diverse class of more than 25000 biomolecules that include familiar compounds such as cholesterol, chlorophyll, vitamin A, ubiquinone, and natural rubber. The two essential building blocks of all isoprenoids, isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), are ubiquitous in the three domains of life. In most eukaryotes and archaea, IPP and DMAPP are generated through the mevalonate pathway. We have identified two novel enzymes, mevalonate-3-kinase and mevalonate-3-phosphate-5-kinase from Thermoplasma acidophilum, which act sequentially in a putative alternate mevalonate pathway. We propose that a yet unidentified ATP-independent decarboxylase acts upon mevalonate 3,5-bisphosphate, yielding isopentenyl phosphate, which is subsequently phosphorylated by the known isopentenyl phosphate kinase from T. acidophilum to generate the universal isoprenoid precursor, IPP.

show abstract

“…It was, therefore, gratifying to find that purified "anhydrodecarboxylase" catalyzed the coordinated removal of the carboxyl group and of the tertiary hydroxy function as postulated (44). Data obtained with 180 suggest that 3-phosphomevalonic-5-pyrophosphate is a transitory intermediate (51), ATP serving as the phosphorylating agent for the tertiary hydroxyl group and thereby promoting its elimination (Fig. 12).…”

Section: Mevalonic Acidmentioning

confidence: 99%

The Biological Synthesis of Cholesterol

Bloch

1965

Science

Self Cite

496

251

View full text Add to dashboard Cite

In the early 1930's after decades of effort the structural elucidation of cholesterol had reached the stage of completion, and with this achievement one of the most brilliant chapters of organic chemistry came to a close. To chemists and biochemists alike the structure of cholesterol at once presented a biosynthetic problem of exceptional challenge. At first sight the molecular architecture of cholesterol seemed enigmatic and devoid of any clues as to how this complex molecule might be constructed from the smaller molecules available in the cell. It is, therefore, all the more remarkable that much of the early speculative thinking came so close to predicting some of the general principles that were eventually shown to operate in sterol biosynthesis. The early proposals were mainly concerned with the problem of ring formation, and quite uniformly they envisioned an origin of the tetracyclic steroidal ring system from an appropriately folded longchain precursor. This intuition proved to be correct. All of the speculative schemes have in some way influenced the research that was to take place later, but none equalled in perspicacity L. Ruzicka's unifying hypothesis on the common origin of terpenes and steroids and the suggestion by Sir Robert Robinson that cholesterol might arise by cyclization of the hydrocarbon squalene. These bold and appealing ideas carried perhaps more weight because they had some experimental evidence to support them. Already in 1926 Shannon, following a proposal by Copyright ? 1965 by the Nobel Foundation.The author is Higgins Professor of Biochemistry at Harvard University, Cambridge, Massachusetts. This article is the lecture he delivered in Stockholm, Sweden, 11 December 1964, when he received the Nobel Prize in medicine and physiology, a prize which he shared with Professor Feodor Lynen of the Max-Planck-Institut fiir Zellchemie, Munich. It is published here with the permission of the Nobel Foundation and will also be included in the complete volumes of Nobel lectures in English, published by the Elsevier Publishing Company, Amsterdam and New York. OCTOBER 1965Heilbron, Kamm, and Owens (1), had shown that squalene fed to animals increases the cholesterol content of the tissues (2).The phase of continuous research on cholesterol biosynthesis begins in 1937 with two independent and remarkably complementary investigations. In the course of their pioneering studies on intermediary metabolism with the aid of stable isotopes, Rittenberg and Schoenheimer at Columbia arrived at the conclusion that the process of cholesterol formation involves the coupling of smaller molecules, "possibly those which have been postulated to be intermediates in the fat and carbohydrate metabolism" (3). During the same year Sonderhoff and Thomas, noting an incorporation of trideuterioacetate into the unsaponifiable materials of yeast, stated: "Es lasst Sich darauf schliessen dass die Sterine der Hefe auf einem recht unmittelbaren Wege aus der Essigsiure entstehen" (4).As a graduate student under Hans T. Clarke at Col...

show abstract

On the Mechanism of Formation of Isopentenylpyrophosphate^*

Cited by 38 publications

References 32 publications

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

Evidence of a Novel Mevalonate Pathway in Archaea

The Biological Synthesis of Cholesterol

Contact Info

Product

Resources

About

On the Mechanism of Formation of Isopentenylpyrophosphate*

Cited by 38 publications

References 32 publications

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

Structural analysis of mevalonate‐3‐kinase provides insight into the mechanisms of isoprenoid pathway decarboxylases

Evidence of a Novel Mevalonate Pathway in Archaea

The Biological Synthesis of Cholesterol

Contact Info

Product

Resources

About

On the Mechanism of Formation of Isopentenylpyrophosphate^*