dc.contributor.author | Bandla, Sravanthi | |
dc.contributor.author | Diaz, Suraya | |
dc.contributor.author | Nasheuer, Heinz-Peter | |
dc.contributor.author | FitzGerald, Una | |
dc.date.accessioned | 2020-01-06T11:57:27Z | |
dc.date.available | 2020-01-06T11:57:27Z | |
dc.date.issued | 2019-08-29 | |
dc.identifier.citation | Bandla, Sravanthi, Diaz, Suraya, Nasheuer, Heinz-Peter, & FitzGerald, Una. (2019). ATPase activity of human binding immunoglobulin protein (BiP) variants is enhanced by signal sequence and physiological concentrations of Mn2+. FEBS Open Bio, 9(8), 1355-1369. doi: 10.1002/2211-5463.12645 | en_IE |
dc.identifier.issn | 2211-5463 | |
dc.identifier.uri | http://hdl.handle.net/10379/15670 | |
dc.description.abstract | B-cell immunoglobulin binding protein (BiP) is an essential endoplasmic reticulum (ER) chaperone normally found in the ER lumen. However, BiP also has other extracellular and intracellular functions. As it is unclear whether peripheral BiP has a signal and/or ER retention sequence, here we produced and biochemically characterised four variants of BiP. The variants differed depending on the presence or the absence of signal and ER retention peptides. Proteins were purified using nickel affinity chromatography, and variant size and quality were confirmed using SDS/PAGE gels. The thermal denaturing temperature of these proteins was found to be 46-47 degrees C. In addition, we characterised nucleotide binding properties in the absence and the presence of divalent cations. Interestingly, in the absence of cations, ADP has a higher binding affinity to BiP than ATP. The presence of divalent cations results in a decrease of the K-d values of both ADP and ATP, indicating higher affinities of both nucleotides for BiP. ATPase assays were carried out to study the enzyme activity of these variants and to characterise the kinetic parameters of BiP variants. Variants with the signal sequence had higher specific activities than those without. Both Mg2+ and Mn2+ efficiently stimulated the ATPase activity of these variants at low micromolar concentrations, whereas calcium failed to stimulate BiP ATPase. Our novel findings indicate the potential functionality of BiP variants that retain a signal sequence, and also reveal the effect of physiological concentrations of cations on the nucleotide binding properties and enzyme activities of all variants. | en_IE |
dc.description.sponsorship | Funding: SB was funded by a Hardiman scholarship of NUI Galway, and SB and UF are supported by the NUIG Foundation Office. SD was funded by the Else Kröner‐Fresenius‐Stiftung (Bad Homburg, Germany) grant no. 2013_A215 to HPN. | en_IE |
dc.format | application/pdf | en_IE |
dc.language.iso | en | en_IE |
dc.publisher | Wiley Open Access | en_IE |
dc.relation.ispartof | FEBS open bio | en |
dc.rights | Attribution-NonCommercial-NoDerivs 3.0 Ireland | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/3.0/ie/ | |
dc.subject | ENDOPLASMIC-RETICULUM | en_IE |
dc.subject | PEPTIDE-BINDING | en_IE |
dc.subject | CHAPERONE BIP | en_IE |
dc.subject | GRP78 | en_IE |
dc.subject | STIMULATION | en_IE |
dc.subject | RELEASE | en_IE |
dc.subject | ANTIGEN | en_IE |
dc.subject | CELLS | en_IE |
dc.subject | SITE | en_IE |
dc.subject | ADP | en_IE |
dc.title | ATPase activity of human binding immunoglobulin protein (BiP) variants is enhanced by signal sequence and physiological concentrations of Mn2+ | en_IE |
dc.type | Article | en_IE |
dc.date.updated | 2020-01-06T10:23:59Z | |
dc.identifier.doi | 10.1002/2211-5463.12645 | |
dc.local.publishedsource | https://doi.org/10.1002/2211-5463.12645 | en_IE |
dc.description.peer-reviewed | peer-reviewed | |
dc.contributor.funder | Hardiman Research Scholarship, National University of Ireland Galway | en_IE |
dc.contributor.funder | Foundation Office, National University of Ireland Galway | en_IE |
dc.contributor.funder | Else Kröner‐Fresenius‐Stiftung | en_IE |
dc.internal.rssid | 17177860 | |
dc.local.contact | Heinz-Peter Nasheuer, 2fn, Rm Bms 2010, Biochemistry S, Bioresearch Building, Nui Galway, Galway. 2430 Email: heinz.nasheuer@nuigalway.ie | |
dc.local.copyrightchecked | Yes | |
dc.local.version | ACCEPTED | |
nui.item.downloads | 229 | |