prikaz prve stranice dokumenta Expression, Purification, and Biochemical Characterization of Human Afamin
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Scientific paper - Original scientific paper
Expression, Purification, and Biochemical Characterization of Human Afamin
Journal of Proteome Research, 17 (2018), 3; 1269-1277. https://doi.org/10.1021/acs.jproteome.7b00867
Altamirano, Alessandra; Naschberger, Andreas; Fürnrohr, Barbara G.; Saldova, Radka; Struwe, Weston B.; Jennings, Patrick M.; Millán Martín, Silvia; Malić, Suzana; Plangger, Immanuel; Lechner, Stefan; Pisano, Reina; Peretti, Nicole; Linke, Bernd; Aguiar, Mario M.; Fresser, Friedrich; Ritsch, Andreas; Lenac Roviš, Tihana; Goode, Christina; Rudd, Pauline M.; Scheffzek, Klaus; Rupp, Bernhard; Dieplinger, Hans More authors...

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Altamirano, A., Naschberger, A., Fürnrohr, B. G., Saldova, R., Struwe, W. B., Jennings, P. M. ... Dieplinger, H. (2018). Expression, Purification, and Biochemical Characterization of Human Afamin. Journal of Proteome Research, 17. (3), 1269-1277. doi: 10.1021/acs.jproteome.7b00867

Altamirano, Alessandra, et al. "Expression, Purification, and Biochemical Characterization of Human Afamin." Journal of Proteome Research, vol. 17, no. 3, 2018, pp. 1269-1277. https://doi.org/10.1021/acs.jproteome.7b00867

Altamirano, Alessandra, Andreas Naschberger, Barbara G. Fürnrohr, Radka Saldova, Weston B. Struwe, Patrick M. Jennings, Silvia Millán Martín, et al. "Expression, Purification, and Biochemical Characterization of Human Afamin." Journal of Proteome Research 17, no. 3 (2018): 1269-1277. https://doi.org/10.1021/acs.jproteome.7b00867

Altamirano, A., et al. (2018) 'Expression, Purification, and Biochemical Characterization of Human Afamin', Journal of Proteome Research, 17(3), pp. 1269-1277. doi: 10.1021/acs.jproteome.7b00867

Altamirano A, Naschberger A, Fürnrohr BG, Saldova R, Struwe WB, Jennings PM, and sur.. Expression, Purification, and Biochemical Characterization of Human Afamin. Journal of Proteome Research [Internet]. 2018 [cited 2025 February 27];17(3):1269-1277. doi: 10.1021/acs.jproteome.7b00867

A. Altamirano, et al., "Expression, Purification, and Biochemical Characterization of Human Afamin", Journal of Proteome Research, vol. 17, no. 3, pp. 1269-1277, 2018. [Online]. Available at: https://urn.nsk.hr/urn:nbn:hr:184:777985. [Accessed: 27 February 2025]

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Metadata
Title (english)Expression, Purification, and Biochemical Characterization of Human Afamin
AuthorAlessandra Altamirano
AuthorAndreas Naschberger
AuthorBarbara G. Fürnrohr
AuthorRadka Saldova
AuthorWeston B. Struwe
AuthorPatrick M. Jennings
AuthorSilvia Millán Martín
AuthorSuzana Malić
AuthorImmanuel Plangger
AuthorStefan Lechner
AuthorReina Pisano
AuthorNicole Peretti
AuthorBernd Linke
AuthorMario M. Aguiar
AuthorFriedrich Fresser
AuthorAndreas Ritsch
AuthorTihana Lenac Roviš
AuthorChristina Goode
AuthorPauline M. Rudd
AuthorKlaus Scheffzek
AuthorBernhard Rupp
AuthorHans Dieplinger
Author's institutionUniversity of Rijeka
Faculty of Medicine
(Center for Proteomics)
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Basic Medical Sciences
Abstract (english)
Afamin is an 87 kDa glycoprotein with five predicted N-glycosylation sites. Afamin's glycan abundance contributes to conformational and chemical inhomogeneity presenting great challenges for molecular structure determination. For the purpose of studying the structure of afamin, various forms of recombinantly expressed human afamin (rhAFM) with different glycosylation patterns were thus created. Wild-type rhAFM and various hypoglycosylated forms were expressed in CHO, CHO-Lec1, and HEK293T cells. Fully nonglycosylated rhAFM was obtained by transfection of point-mutated cDNA to delete all N-glycosylation sites of afamin. Wild-type and hypo/nonglycosylated rhAFM were purified from cell culture supernatants by immobilized metal ion affinity and size exclusion chromatography. Glycan analysis of purified proteins demonstrated differences in micro- and macro-heterogeneity of glycosylation enabling the comparison between hypoglycosylated, wild- type rhAFM, and native plasma afamin. Because antibody fragments can work as artificial chaperones by stabilizing the structure of proteins and consequently enhance the chance for successful crystallization, we incubated a Fab fragment of the monoclonal anti-afamin antibody N14 with human afamin and obtained a stoichiometric complex. Subsequent results showed sufficient expression of various partially or nonglycosylated forms of rhAFM in HEK293T and CHO cells and revealed that glycosylation is not necessary for expression and secretion
Keywords (english)
Languageenglish
Publication typeScientific paper - Original scientific paper
Publication statusPublished
Peer reviewPeer review - international
Publication versionPublished version
Journal titleJournal of Proteome Research
Numberingvol. 17, no. 3, pp. 1269-1277
p-ISSN1535-3893
DOIhttps://doi.org/10.1021/acs.jproteome.7b00867
URN:NBNurn:nbn:hr:184:777985
Publication2018
Document URLhttp://bib.irb.hr/1049761
Type of resourceText
Access conditionsClosed access
Terms of useLicence In copyright icon
Created on2022-02-21 11:58:42