Crystal structures of native and recombinant yeast fumarase 1 1Edited by D. Rees

Weaver, Todd; Lees, M.; Zaĭtsev, V. N.; Zaitseva, I. Ya.; Duke, Elizabeth; Lindley, P.L; McSweeny, S.; Svensson, Anders; Keruchenko, Jan S.; Keruchenko, Irina D.; Gladilin, K.L.; Banaszak, Leonard J.

doi:10.1006/jmbi.1998.1862

Cited by 51 publications

(51 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1, substitutions of five amino acids (M5-SALVA) of yeast fumarase for amino acids frequently found at corresponding positions in bacterial fumarases still abolished dual targeting. The second sequence examined was via a deletion of a 16 amino acid sequence of fumarase that has been implicated as being part of the active site of the enzyme (24). Like the other deletions, this deletion caused a total mis-distribution of mature fumarase to mitochondria (Fig.…”

Section: Subcellular Localization Of Mutant Fumarases-mentioning

confidence: 99%

“…Interestingly, the deletion in ⌬〉 occurs within the middle domain, whereas the other deletions occur within the aminoand carboxyl-terminal domains. In fact, the crystal structure of fumarase (24) indicates that the middle domain forms a fivehelix bundle which, in the full protein, together with the middle domains of the other three subunits, forms a bundle of 20 ␣-helices, which, in turn, probably hold the tetramer together. With respect to subcellular distribution, formation of the tetramer does not ensure that dual targeting will occur.…”

Section: Subcellular Localization Of Mutant Fumarases-mentioning

confidence: 99%

See 1 more Smart Citation

Folding of Fumarase during Mitochondrial Import Determines its Dual Targeting in Yeast

Sass¹,

Karniely²,

Pines

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

We have previously proposed that a single translation product of the FUM1 gene encoding fumarase is distributed between the cytosol and mitochondria of Saccharomyces cerevisiae and that all fumarase translation products are targeted and processed in mitochondria before distribution. Thus, fumarase processed in mitochondria returns to the cytosol. In the current work, we (i) generated mutations throughout the coding sequence which resulted in fumarases with altered conformations that are targeted to mitochondria but have lost their ability to be distributed; (ii) showed by mass spectrometry that mature cytosolic and mitochondrial fumarase isoenzymes are identical; and (iii) showed that hsp70 chaperones in the cytosol (Ssa) and mitochondria (Ssc1) can affect fumarase distribution. The results are discussed in light of our model of targeting and distribution, which suggests that rapid folding of fumarase into an import-incompetent state provides the driving force for retrograde movement of the processed protein back to the cytosol through the translocation pore.

show abstract

Section: Subcellular Localization Of Mutant Fumarases-mentioning

confidence: 99%

Section: Subcellular Localization Of Mutant Fumarases-mentioning

confidence: 99%

Folding of Fumarase during Mitochondrial Import Determines its Dual Targeting in Yeast

Sass¹,

Karniely²,

Pines

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Many organisms have genes corresponding to both the classes, as in Escherichia coli (Ec), which has three FH encoding genes viz., fum A, B and C. Fum A and fum B are 4Fe-4S cluster containing class I enzymes, while Fum C belongs to class II type FH. Recently, another gene fum D has been identified in the E. coli genome to code for a class I fumarase with altered substrate preferences (14).The structural and biochemical characteristics of class II FH are thoroughly studied from different organisms viz., human, porcine, yeast, E. coli and other sources (15)(16)(17)(18)(19). On the other hand, class I FH is not well studied owing to its thermolabile and oxygen sensitive nature.…”

mentioning

confidence: 99%

Biochemical characterization and essentiality of fumarate hydratase

Jayaraman¹,

Suryavanshi²,

Kalale

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Plasmodium falciparum (Pf), the causative agent of malaria, has an iron-sulfur cluster-containing class I fumarate hydratase (FH) that catalyzes the interconversion of fumarate to malate, a wellknown reaction in the tricarboxylic acid cycle. In humans, the same reaction is catalyzed by class II FH that has no sequence or structural homology with the class I enzyme from Plasmodium. Fumarate is generated in large quantities in the parasite as a byproduct of AMP synthesis and is converted to malate by FH and then used in the generation of the key metabolites oxaloacetate, aspartate, and pyruvate. Previous studies have identified the FH reaction as being essential to P. falciparum, but biochemical characterizations of PfFH that may provide leads for the development of specific inhibitors are lacking. Here, we report on the kinetic characterization of purified recombinant PfFH, functional complementation of fh deficiency in Escherichia coli, and mitochondrial localization in the parasite. We found that the substrate analog mercaptosuccinic acid is a potent PfFH inhibitor, with a K i value in the nanomolar range. The fh gene could not be knocked out in Plasmodium berghei when transfectants were introduced into BALB/c mice; however, fh knockout was successful when C57BL/6 mice were used as host, suggesting that the essentiality of the fh gene to the parasite was mouse strain dependent.Plasmodium falciparum (Pf), the causative agent of the most lethal form of malaria, during its intra-erythrocytic asexual stages, derives ATP primarily from glycolysis with low contribution from mitochondrial pathways (1, 2). The bulk of pyruvate formed is converted to lactic acid with a minor amount entering the tricarboxylic acid (TCA) cycle, the flux through which is upregulated in sexual stages (2). Key intermediates that anaplerotically feed into the TCA cycle are α-ketoglutarate derived from glutamate, oxaloacetate (OAA) from phosphoenolpyruvate, and fumarate from adenosine 5΄-monophosphate (AMP) synthesis. Synthesis of AMP in the parasite is solely from inosine-5΄-monophosphate (IMP) through a pathway involving the enzymes adenylosuccinate synthetase (ADSS) and adenylosuccinate lyase (ASL). The net reaction of ADSS and ASL involves consumption of GTP and aspartate and, generation of GDP, P i and fumarate. In the rapidly dividing parasite with an AT-rich genome and high energy requirements, leading to a high demand for adenine pools, one would expect a high flux of fumarate generation. The parasite does not secrete fumarate but instead, the carbon derived from this metabolite can be traced in malate, OAA, aspartate, pyruvate (through PEP) and lactate (3). The metabolic significance of this fumarate anaplerosis is still obscure. In this context, fumarate hydratase (FH, fumarase) the key enzyme to metabolize fumarate becomes an Fumarate hydratase (fumarase, E.C. 4.2.1.2) catalyzes the reversible conversion of fumarate to malate. The stereospecific reaction involves the anti-addition of a water molecule across the carbon-carbon...

show abstract

“…The crystal structures of the E. coli fumarase C and Saccharomyces cerevisiae fumarase have been elucidated. 9,10 The high degree of homology of these Supported by Cancer Research UK (clinical research fellowship to N.A.A. ).…”

mentioning

confidence: 99%

“…The crystal structures of the E. coli fumarase C and Saccharomyces cerevisiae fumarase have been elucidated. 9,10 The high degree of homology of these proteins to human FH allows their use as models for predicting the effect of missense mutations on FH function. The FH protein exists as a homotetramer with two substrate-binding sites (Figure 1), designated the A-site and the B-site.…”

mentioning

confidence: 99%

Missense Mutations in Fumarate Hydratase in Multiple Cutaneous and Uterine Leiomyomatosis and Renal Cell Cancer

Alam

Olpin

Rowan

et al. 2005

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

Heterozygous germline mutations in fumarate hydratase (FH) predispose to the multiple cutaneous and uterine leiomyomatosis syndrome (MCUL), which, when co-existing with renal cancer, is also known as hereditary leiomyomatosis and renal cell cancer. Twenty-seven distinct missense mutations represent 68% of FH mutations reported in MCUL. Here we show that FH missense mutations significantly occurred in fully conserved residues and in residues functioning in the FH A-site, B-site, or subunit-interacting region. Of 24 distinct missense mutations, 13 (54%) occurred in the substrate-binding Asite, 4 (17%) in the substrate-binding B-site, and 7 (29%) in the subunit-interacting region. Clustering of missense mutations suggested the presence of possible mutational hotspots. FH functional assay of lymphoblastoid cell lines from 23 individuals with heterozygous FH missense mutations showed that A-site mutants had significantly less residual activity than B-site mutants, supporting data from Escherichia coli that the A-site is the main catalytic site. Missense FH mutations predisposing to renal cancer had no unusual features, and identical mutations were found in families without renal cancer, suggesting a role for genetic or environmental factors in renal cancer development In the autosomal dominant syndrome of multiple cutaneous and uterine leiomyomatosis (MCUL, Reed syndrome, leiomyomatosis cutis et uteri, multiple leiomyomatosis; OMIM 150800), affected females develop uterine leiomyomas and affected individuals of both sexes develop cutaneous leiomyomas. 1 Cutaneous leiomyomas are believed to be derived from the smooth muscle of the pilo-arrector apparatus. They generally present in the second, third, or fourth decades, typically as grouped papules or nodules on the trunk or limbs and are characteristically painful particularly in response to low temperatures or touch. Uterine leiomyomas or fibroids in MCUL are severely symptomatic with a large proportion of patients requiring symptom control by hysterectomy. 2 A small proportion of families with MCUL also cluster renal cancer, either papillary renal type II cancer or renal collecting duct cancer. 1,[3][4][5][6] This disease variant has been referred to as hereditary leiomyomatosis and renal cancer (HLRCC, OMIM 605839). MCUL/HLRCC has been found to be caused by germline mutations in fumarate hydratase (FH) in the majority of screened cases. 1,5,6 Forty-six distinct FH mutations have been reported to date in MCUL/HLRCC. Twenty-seven of these are missense mutations of 26 different residues (one residue has two reported mutations, R190H and R190L). These 27 distinct missense mutations represent 55 of 81 (68%) of the FH mutations reported in MCUL probands. 1,[5][6][7] The FH locus encodes two isoforms of fumarate hydratase, cytosolic and mitochondrial, which differ only in that the latter has an initial mitochondrial signal peptide. Fumarate hydratase catalyzes the stereospecific reversible hydration of fumarate to L-malate. The mitochondrial isoform performs this reactio...

show abstract

Crystal structures of native and recombinant yeast fumarase 1 1Edited by D. Rees

Cited by 51 publications

References 21 publications

Folding of Fumarase during Mitochondrial Import Determines its Dual Targeting in Yeast

Folding of Fumarase during Mitochondrial Import Determines its Dual Targeting in Yeast

Biochemical characterization and essentiality of fumarate hydratase

Missense Mutations in Fumarate Hydratase in Multiple Cutaneous and Uterine Leiomyomatosis and Renal Cell Cancer

Contact Info

Product

Resources

About