Refined Crystal Structures of Native Human Angiogenin and Two Active Site Variants: Implications for the Unique Functional Properties of an Enzyme Involved in Neovascularisation During Tumour Growth

Leonidas, D.D.; Shapiro, Robert; Allen, S.C.; Subbarao, G.V.; Veluraja, K.; Acharya, K. Ravi

doi:10.1006/jmbi.1998.2378

Cited by 94 publications

(170 citation statements)

References 112 publications

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“…Angiogenin was claimed to specifically cleave tRNAs. But, neither a specific cleavage site nor a specific tRNA species as a substrate has been reported (28), which is in clear contrast to colicins D and E5.…”

Section: Discussionmentioning

confidence: 87%

A cytotoxic ribonuclease which specifically cleaves four isoaccepting arginine tRNAs at their anticodon loops

Tomita

Ogawa

Uozumi

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

158

143

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Colicin D has long been thought to stop protein synthesis in infected Escherichia coli cells by inactivating ribosomes, just like colicin E3. Here, we show that colicin D specifically cleaves tRNAs Arg including four isoaccepting molecules both in vivo and in vitro. The cleavage occurs in vitro between positions 38 and 39 in an anticodon loop with a 2 ,3 -cyclic phosphate end, and is inhibited by a specific immunity protein. Consistent with the cleavage of tRNAs Arg , the RNA fraction of colicin-treated cells significantly reduced the amino acid-accepting activity only for arginine. Furthermore, we generated a single mutation of histidine in the C-terminal possible catalytic domain, which caused the loss of the killing activity in vivo together with the tRNA Arg -cleaving activity both in vivo and in vitro. These findings show that colicin D directly cleaves cytoplasmic tRNAs Arg , which leads to impairment of protein synthesis and cell death. Recently, we found that colicin E5 stops protein synthesis by cleaving the anticodons of specific tRNAs for Tyr, His, Asn, and Asp. Despite these apparently similar actions on tRNAs and cells, colicins D and E5 not only exhibit no sequence homology but also have different molecular mechanisms as to both substrate recognition and catalytic reaction.C olicins are plasmid-encoded proteins that are toxic to Escherichia coli cells that do not have the same plasmid or a cognate Col plasmid (1-5). Most colicins are produced in response to SOS-inducing signals and are secreted into the medium. After binding to cell-surface receptors on sensitive cells, they are translocated across the membrane and then exert their final cytotoxic activities, which are attributable to their C-terminal domains. Two major modes of toxicity are well known; colicins A, B, E1, Ia, Ib, K, and N are ion-channel formers attacking the cytoplasmic membrane, and colicins E2 to E9 are nucleases. In the latter group, E2, E7, E8, and E9 are DNases, and E3 is a special kind of RNase that cleaves 16S rRNA within ribosomes.Colicins E4 to E6 quickly stop amino acid incorporation in treated cells (6), suggesting impairment of protein synthesis analogous in the established case of E3, which specifically cleaves 16S-RNA at the 49th bond from the 3Ј end leading to inactivation of ribosomes (7-9). This is also the case with colicin D, which Timmis and Hedges have characterized as to the physiological response of treated cells (10, 11), although the actual molecular basis of the cytotoxic effect of colicin D, as well as those of E4 to E6, remained to be elucidated. Colicins E4 and E6 proved to be E3-homologs and showed comparable activity toward ribosomes (ref. 12; GenBank accession number X63621; Y. Gunji, M. Ohno, T.O., H.M., and T.U., unpublished data), but colicin E5 exhibits no similarity to E3 in the C-terminal active domain. We recently showed that colicin E5 comprises a third category of nuclease-type colicins, which does not attack ribosomes but specific tRNAs (13). E5 is a novel RNase that cleaves the anticodons...

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Section: Discussionmentioning

confidence: 87%

A cytotoxic ribonuclease which specifically cleaves four isoaccepting arginine tRNAs at their anticodon loops

Tomita

Ogawa

Uozumi

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

158

143

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“…4 and 11). The importance of such selective amino acid residue mediated path for different mutants is supported by published crystal structure of ANG, which established Leu115 as a factor responsible for weak ribonucleolytic activity [21]. Mutants V113I, R121C, R121H, K54E, F100I, A98V and P123L, which do not show any conformational switching of His114, exhibited no hydrogen bond interaction path from the site of mutation to His114.…”

Section: Resultsmentioning

confidence: 94%

“…Crystal structure of human angiogenin (PDB: 1B1I) was used as the starting structure and desired residues were mutated in silico keeping its secondary structure intact [21]. Heteroatoms, cofactor CIT and crystallographic water were removed a priori.…”

Section: Generalized Born Solvent Molecular Simulationsmentioning

confidence: 99%

Fast prediction of deleterious angiogenin mutations causing amyotrophic lateral sclerosis

et al. 2013

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a b s t r a c tCertain single nucleotide polymorphisms causing missense mutations in angiogenin result in its loss-of-function and onset of amyotrophic lateral sclerosis (ALS). Although several such associations are reported across diverse ethnic groups, no method is available for predicting if a new mutation is deleterious. We present here a fast molecular dynamics based method for determining the mechanisms of functional loss caused by mutations, and attributes to ascertain whether a mutation causes ALS.

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“…Crystal structures of ANG show that Gln12 forms part of the P 1 subsite where phosphodiester bond cleavage occurs; it makes hydrogen bonds with Lys40 (32) and, in the complex of ANG with phosphate, with ligand as well (46). However, previous studies on this residue (47) and on its counterpart Gln11 in RNase A (48) have shown that replacement by Ala does not cause any significant loss of activity, which therefore indicates that Gln12/Gln11 is not a critical residue.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Human Angiogenin Variants Implicated in Amyotrophic Lateral Sclerosis

et al. 2007

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Human angiogenin (ANG), the first member of the angiogenin family (from the pancreatic ribonuclease A superfamily) to be identified, is an angiogenic factor that induces neovascularization. It has received much attention due to its involvement in the growth of tumors and its elevated expression level in pancreatic and several other cancers. Recently the biological role of ANG has been shown to extend to the nervous system. Mutations in ANG have been linked with familial as well as sporadic forms of amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disorder characterized by selective destruction of motor neurons. Furthermore, mouse angiogenin-1 has been shown to be expressed in the developing nervous system and during the neuronal differentiation of pluripotent stem cells. We have now characterized the seven variants of ANG reported in ALS patients with respect to the known biochemical properties of ANG and further studied the biological properties of three of these variants. Our results show that the ribonucleolytic activity of six of the seven ANG-ALS implicated variants is significantly reduced or lost and some variants also show altered thermal stability. We report a significant reduction in the cell proliferative and angiogenic activities of the three variants that we chose to investigate further. Our studies on the biochemical and structural features of these ANG variants now form the basis for further investigations to determine their role(s) in ALS.Amyotrophic lateral sclerosis (ALS) 1 is a fatal neurodegenerative disorder characterized by selective destruction of motor neurons (1). In the past decade, a small number of genes involved in the etiology of the disease have been identified (for a recent review see ref 2). The best studied of these is SOD1 (3), the gene for Cu/Zn superoxide dismutase. More than a hundred SOD1 mutations have now been linked with ALS, and motor neuron death from many of these mutations has been shown to result from a toxic gain of function rather than loss of dismutase activity. However, mutations in SOD1 account for only 1-2% of all cases of ALS and 20% of the familial cases. Some of the other proteins implicated in ALS are the vesicle-trafficking protein VAPB (4); ALSIN, a putative guanine nucleotide factor for GTPase (5); and senataxin (6). In addition, vascular endothelial growth factor (VEGF), an angiogenic factor that plays an important role in motor neuron survival, has been linked with ALS (7-10). However, the causes and molecular mechanisms underlying ALS are still largely unclear, and effective therapies do not appear to be imminent. Angiogenin (ANG), which encodes an angiogenic protein, was recently identified as a candidate susceptibility gene for ALS in the Irish and Scottish population by Greenway et al. (11). In a further study of over 2500 individuals from five independent populations from northern Europe and North America, Greenway et al. (12) found seven missense mutations (Figure 1) in 11 unrelated individuals with sporadic ALS and in f...

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Refined Crystal Structures of Native Human Angiogenin and Two Active Site Variants: Implications for the Unique Functional Properties of an Enzyme Involved in Neovascularisation During Tumour Growth

Cited by 94 publications

References 112 publications

A cytotoxic ribonuclease which specifically cleaves four isoaccepting arginine tRNAs at their anticodon loops

A cytotoxic ribonuclease which specifically cleaves four isoaccepting arginine tRNAs at their anticodon loops

Fast prediction of deleterious angiogenin mutations causing amyotrophic lateral sclerosis

Characterization of Human Angiogenin Variants Implicated in Amyotrophic Lateral Sclerosis

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