The Coding ECP 434(G&amp;gt;C) Gene Polymorphism Determines the Cytotoxicity of ECP but Has Minor Effects on Fibroblast-Mediated Gel Contraction and No Effect on RNase Activity

Rubin, Jenny; Zagai, Ulrika; Blom, Kristin; Trulson, Agneta; Engström, Åke

doi:10.4049/jimmunol.0803912

Cited by 27 publications

(29 citation statements)

References 35 publications

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“…Site-directed mutagenesis studies identifi ed the main critical residues exposed at the protein surface involved in membrane lysis and cytotoxicity (Carreras et al , 2003 ;Torrent et al , 2007 ;Singh and Batra , 2011 ). Additionally, complementary studies on the protein native posttranslational forms and comparison of polymorphism variants highlighted the contribution of glycosylation on the protein cytotoxic power (Trulson et al , 2007 ;Rubin et al , 2009 ;Woschnagg et al , 2009 ). In the following section, we detail the current knowledge on ECP antimicrobial properties.…”

Section: The Eosinophil Cationic Protein (Ecp) In Host Defencementioning

confidence: 99%

Structural determinants of the eosinophil cationic protein antimicrobial activity

Boix

Salazar

Torrent

et al. 2012

Biological Chemistry

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Antimicrobial RNases are small cationic proteins belonging to the vertebrate RNase A superfamily and endowed with a wide range of antipathogen activities. Vertebrate RNases, while sharing the active site architecture, are found to display a variety of noncatalytical biological properties, providing an excellent example of multitask proteins. The antibacterial activity of distant related RNases suggested that the family evolved from an ancestral host-defence function. The review provides a structural insight into antimicrobial RNases, taking as a reference the human RNase 3, also named eosinophil cationic protein (ECP). A particular high binding affi nity against bacterial wall structures mediates the protein action. In particular, the interaction with the lipopolysaccharides at the Gram-negative outer membrane correlates with the protein antimicrobial and specifi c cell agglutinating activity. Although a direct mechanical action at the bacteria wall seems to be suffi cient to trigger bacterial death, a potential intracellular target cannot be discarded. Indeed, the cationic clusters at the protein surface may serve both to interact with nucleic acids and cell surface heterosaccharides. Sequence determinants for ECP activity were screened by prediction tools, proteolysis and peptide synthesis. Docking results are complementing the structural analysis to delineate the protein anchoring sites for anionic targets of biological signifi cance.

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Section: The Eosinophil Cationic Protein (Ecp) In Host Defencementioning

confidence: 99%

Structural determinants of the eosinophil cationic protein antimicrobial activity

Boix

Salazar

Torrent

et al. 2012

Biological Chemistry

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“…Eosinophil samples (each containing 10 ng of ECP) or 0.5 g of purified ECP (see Ref. 16 for details on ECP purification) were added to the wells and the arrays were incubated in 4°C overnight with mild shaking.…”

Section: Seldi-tof Ms Affinity Capture Assaymentioning

confidence: 99%

“…ECP (HMW-ECP) and one low m.w. ECP (LMW-ECP) pool (16). The two ECP pools were characterized by SELDI-TOF MS. Fig.…”

Section: Cytotoxicity Of Purified Ecpmentioning

confidence: 99%

Eosinophil Cationic Protein (ECP) Is Processed during Secretion

Woschnagg

Rubin

Venge

2009

The Journal of Immunology

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The eosinophil granulocyte is an inflammatory cell involved in allergic diseases such as asthma and rhinitis. Eosinophil cationic protein (ECP) is a basic and potentially cytotoxic granule protein that is released from the eosinophil upon activation. The aim was to study secretion of molecular variants of ECP from blood eosinophils with the hypothesis that the stored noncytotoxic ECP is altered into cytotoxic species upon release from the cell. Eosinophil granulocytes were purified to >95% from venous blood from birch pollen allergic subjects, with symptoms of rhinitis, and from healthy control subjects during the birch pollen season. The cells were stimulated with IL-5, GM-CSF, or serum-opsonized Sephadex particles. Concentration of ECP in cells or supernatants was measured by means of a fluoroenzyme immunoassay, and ECP heterogeneity was studied using an affinity capture assay with the surface-enhanced laser desorption/ionization-time of flight mass spectrometry technique. Extracts of unstimulated eosinophils contained 10 major ECP variants, with molecular masses ranging from 16.1 to 17.7 kDa. Stimulation with particles mainly induced the secretion of two molecular variants at 16.1 and 16.3 kDa, while cytokine stimulation gave rise to a different secretion profile. ECP variants in the pellet extracts remained unaffected by cell activation. The modifications of secreted ECP were partly explained by differences in N-linked glycosylations. Secretion of ECP from eosinophils involves protein modification. The molecular masses of released ECP have acquired the masses of the cytotoxic species.

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“…Due to G>C sub sti tu tion, the co don for the ami no acid ar gi ni ne is chan ged in the co don for threo ni ne, whi ch lea ds to the chan ge in Ar g97Thr in the ECP pro tein. The consequen ce of the chan ge in ami no acid struc tu re is al te ra tion of bio lo gi cal ac ti vi ty of the ECP mo le cule be cau se Ar g97 is im por ta nt for the cyto toxic acti vi ty of ECP (19,20). The sub sti tu tion of ar gi ni ne at po si tion 97 wi th threo ni ne al lows the for ma tion of a new, four th glyco syla tion si te at po si tion 95 (EC-P 95Asn ), and en han ced glyco syla tion of ECP pro tein dec rea ses its cyto toxic ac ti vi ty.…”

Section: Rna Se3 Po Lymor Phis Msmentioning

confidence: 99%

“…The sub sti tu tion of ar gi ni ne at po si tion 97 wi th threo ni ne al lows the for ma tion of a new, four th glyco syla tion si te at po si tion 95 (EC-P 95Asn ), and en han ced glyco syla tion of ECP pro tein dec rea ses its cyto toxic ac ti vi ty. Stu dies of re combi na nt ECP 97Thr have showed that, af ter deg lycosyla tion, it be co mes cyto toxi cal ly ac ti ve li ke EC-P 97Arg (20). Pa ra si ta ry in fec tio ns, as thma and ma ligna nt di sea ses are on ly so me pat ho lo gi cal con di tions wi th ECP in vol ve me nt in their pat ho mec ha nisms so that di sea se out co me is as so cia ted wi th ECP phe no type or ge no type.…”

Section: Rna Se3 Po Lymor Phis Msmentioning

confidence: 99%

Eosinophil catonic protein – current concepts and controversies

Topić

Dodig

2011

Biochem Med

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Eo si nop hil ca tio nic pro tein (ECP) is a he te ro ge neous mo le cu le ori gi na ti ng from ac ti va ted eo si nop hil gra nu lo cytes. Bio lo gi cal ac ti vi ty and the cel lu lar con te nt of ECP are de ter mi ned by ge ne tic and pos ttran sla tio nal fac to rs. Se ve ral sin gle nuc leo ti de po lymor phis ms (SNPs) in hu man ECP ge ne (RNA-SE3) ha ve been des cri bed so far. ECP is a me dia tor in ho st im mu ne res pon se to pa ra si tes, bac te ria and vi ru ses. By its cyto toxic and no n-cyto toxic ac ti vi ty, ECP may al so cau se si de-eff ec ts in the hos t's own tis sues. The lar ge st num ber of cli ni cal stu dies is fo cu sed on the ro le of ECP in eo si nop hi lre la ted di sor de rs, par ti cu lar ly in as thma. Al thou gh pre se nt in nu me rous bo dy fl ui ds, diffi cu lt bioa vai la bi li ty of bio lo gi cal ma te rial, in va si ve sam pli ng met ho ds and com plex sam ple ma na ge me nt prior to ECP le vel de ter mi na tion are the rea so ns that se rum is mo st com mon ly used in rou ti ne la bo rato ry prac ti ce. As nu me rous bio lo gi cal and met ho do lo gi cal prea na lyti cal fac to rs (the type of col lec tion te st-tu be, tem pe ra tu re and du ra tion of blood clot ti ng, cen tri fu ga tion, he mo lysis) may aff e ct te st re su lt, the sam ple for se rum ECP de ter mi na tion shou ld be col lec ted un der stan dar di zed con di tions. Re gar di ng in ter pre ta tion of re sul ts, it is ne ces sa ry, alo ng wi th ab so lu te ECP con cen tra tion va lues, to mo ni tor chan ges in ECP con cen tra tion du ri ng the du ra tion of di sea se or af ter im ple men ted the ra py, and in ter pret ECP te st re su lt in com bi na tion wi th ot her la bo ra to ry and cli ni cal fi n din gs. Ratio nal ap proa ch to se lec tion of new tes ts is in deed one of im por ta nt requi re men ts that me di cal wor ke rs meet to day. To enab le them to de ter mi ne the cli ni cal sig ni fi can ce of ECP wi th bet ter cer tain ty, fur ther stu dies on a lar ge num ber of spe ci fi c pa tie nt grou ps are nee ded. Key wor ds: eo si nop hil ca tio nic pro tein; po lymor phi sm; cyto toxi ci ty; as thma Eo si nop hil ca tio nic pro tein -cur re nt con cep ts and con tro ver siesRe na ta Zrin ski To pic*, Sla vi ca Do digDe par tme nt of Cli ni cal La bo ra to ry Diag no sis, Sreb r njak Chil dre n's Hos pi tal, Zag reb, Croa tia *Cor res pon di ng aut hor: re na ta.zrinski-topic@zg.t-com.hr Review In tro duc tionEo si nop hil ca tio nic pro tein (ECP), al so known under the na me ri bo nuc lea se 3 (RNa se 3), is a sin gle ca tio nic po lypep ti de chain con sis ti ng of 133 amino aci ds, wi th isoe lec tric poi nt bei ng 10.8 due to a hi gh con te nt of ba sic ami no aci ds, par ti cu lar ly argi ni ne. In the pro tein struc tu re, the re are three poten tial si tes for N-lin ked glyco syla tion: ami no aci ds 57-59, 65-67, and 92-94. Due to the diff e re nt con tent of car bo hydra te su bu ni ts: sia lic acid, ga lac to se and ace tylglu co sa mi ne, the mo le cu lar ma ss of ECP pro tein ra...

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The Coding ECP 434(G>C) Gene Polymorphism Determines the Cytotoxicity of ECP but Has Minor Effects on Fibroblast-Mediated Gel Contraction and No Effect on RNase Activity

Cited by 27 publications

References 35 publications

Structural determinants of the eosinophil cationic protein antimicrobial activity

Structural determinants of the eosinophil cationic protein antimicrobial activity

Eosinophil Cationic Protein (ECP) Is Processed during Secretion

Eosinophil catonic protein – current concepts and controversies

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