KfrA plasmid protein monolayers on latex particles-electrokinetic measurements

Marta Kujda , Zbigniew Adamczyk , Grażyna Jagura-Burdzy , Małgorzata Adamczyk

Abstract

Monolayers of KfrA, a protein assisting in bacteria plasmid segregation, on polystyrene latex particles were produced in controlled self-assembling under diffusion-controlled conditions. The coverage of the protein was quantitatively determined as a function of ionic strength (up to 0.15 M, NaCl) via micro-electrophoretic measurements and concentration depletion with the aid of AFM imaging. The maximum monolayer coverage of KfrA monotonically increased with ionic strength attaining 2.0 mg m−2 for 0.15 M, NaCl that corresponds to the dimensionless coverage of 0.48. This is in accordance with theoretical calculations derived from the random sequential adsorption modeling assuming a tetrameric aggregation state. A high stability of the monolayers in pH cycling experiments was confirmed, which proves the irreversibility of protein adsorption on latex. The acid base properties and the electrokinetic charge of monolayers were also determined via the electrophoretic mobility measurements carried out for various ionic strength. In this way the isoelectric point of the protein of 4.8 was determined, which is prohibitive via bulk measurements. It was concluded that the procedure used in our work is reliable and efficient for characterizing physicochemical properties if minor amounts of a protein are available.
Author Marta Kujda - [Institute of Catalysis and Surface Chemistry of the Polish Academy of Sciences]
Marta Kujda,,
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, Zbigniew Adamczyk - [Institute of Catalysis and Surface Chemistry of the Polish Academy of Sciences]
Zbigniew Adamczyk,,
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, Grażyna Jagura-Burdzy - [Institute of Biochemistry and Biophysics of the Polish Academy of Sciences]
Grażyna Jagura-Burdzy,,
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, Małgorzata Adamczyk (FC / IBC / DDTB)
Małgorzata Adamczyk,,
- Department Of Drug Technology And Biotechnology
Journal seriesColloids and Surfaces B-Biointerfaces, ISSN 0927-7765
Issue year2013
Vol112
Pages165-170
Publication size in sheets0.5
Keywords in EnglishMonolayers of KfrA protein on latex; Plasmid protein adsorption on latex; KfrA plasmid protein properties; Isoelectric point of KfrA; KfrA plasmid protein adsorption; Latex covered by KfrA plasmid protein
ASJC Classification1505 Colloid and Surface Chemistry; 1606 Physical and Theoretical Chemistry; 3110 Surfaces and Interfaces; 2700 General Medicine; 1305 Biotechnology
DOIDOI:10.1016/j.colsurfb.2013.07.026
URL http://www.sciencedirect.com/science/article/pii/S0927776513004621
Languageen angielski
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1-s2.0-S0927776513004621-main.pdf 473.53 KB
Score (nominal)35
Score sourcejournalList
ScoreMinisterial score = 30.0, 02-02-2020, ArticleFromJournal
Ministerial score (2013-2016) = 35.0, 02-02-2020, ArticleFromJournal
Publication indicators Scopus Citations = 6; WoS Citations = 7; Scopus SNIP (Source Normalised Impact per Paper): 2013 = 1.573; WoS Impact Factor: 2013 = 4.287 (2) - 2013=4.226 (5)
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