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The influence of fatty acids on determination of human serum albumin thiol group

Authorized Users Only
2014
Authors
Jovanović, Vesna B.
Pavićević, Ivan D.
Takić, Marija M.
Penezic-Romanjuk, Ana Z.
Aćimović, Jelena M.
Mandić, Ljuba M.
Article (Published version)
Metadata
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Abstract
During investigation of the changes of the Cys34 thiol group of human serum albumin (HSA) (isolated by affinity chromatography with Cibacron Blue (CB)) in diabetes, we found that the HSA-SH content was higher (11-33%) than the total serum thiol content. The influence of fatty acids (FA) binding to HSA on this discrepancy was investigated in vitro (using fluorescence and CD spectroscopy and GC) and with HSA samples from diabetic (n=20) and control groups (n=17). HSA-bound FA determine the selection of HSA molecules by CB and enhance reactivity and/or accessibility of the SH group. A high content of polyunsaturated FA (35.6%) leads to weaker binding of HSA molecules to CB. Rate constants of DTNB reaction with the SH group of HSA applied to a CB column, bound-HSA and unbound-HSA fractions, were 4.8 x 10(-3), 21.6 x 10(-3), and 11.2 x 10(-3) s(-1), respectively. The HSA-SH group of diabetics is more reactive compared with control individuals (rate constants 20.9 x 10(-3)+/- 4.4 x 10(-3) vs... 12.9 x 10(-3)+/- 2.6 x 10(-3) s(-1), P lt 0.05). Recovery values of the SH group obtained after chromatography of HSA with bound stearic acid ranged from 110 to 140%, while those for defatted HSA were from 98.5 to 101.7%. Thus, HSA-bound FA leads to an increase of HSA-SH content and a contribution to total serum thiols, which make the determination of the thiol group unreliable.

Keywords:
HSA-thiol group determination / Fatty acid / Albumin isolation / Affinity chromatography / Diabetes
Source:
Analytical Biochemistry, 2014, 448, 50-57
Publisher:
  • Academic Press Inc Elsevier Science, San Diego
Funding / projects:
  • Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-172049)
  • European Commission, EU Seventh Framework Programme (FP7), RegPot FCUB-ERA, GA [256715]

DOI: 10.1016/j.ab.2013.11.030

ISSN: 0003-2697

PubMed: 24316317

WoS: 000331678500008

Scopus: 2-s2.0-84891671605
[ Google Scholar ]
14
11
URI
http://rimi.imi.bg.ac.rs/handle/123456789/558
Collections
  • Radovi istraživača / Researchers' publications
Institution/Community
Institut za medicinska istraživanja
TY  - JOUR
AU  - Jovanović, Vesna B.
AU  - Pavićević, Ivan D.
AU  - Takić, Marija M.
AU  - Penezic-Romanjuk, Ana Z.
AU  - Aćimović, Jelena M.
AU  - Mandić, Ljuba M.
PY  - 2014
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/558
AB  - During investigation of the changes of the Cys34 thiol group of human serum albumin (HSA) (isolated by affinity chromatography with Cibacron Blue (CB)) in diabetes, we found that the HSA-SH content was higher (11-33%) than the total serum thiol content. The influence of fatty acids (FA) binding to HSA on this discrepancy was investigated in vitro (using fluorescence and CD spectroscopy and GC) and with HSA samples from diabetic (n=20) and control groups (n=17). HSA-bound FA determine the selection of HSA molecules by CB and enhance reactivity and/or accessibility of the SH group. A high content of polyunsaturated FA (35.6%) leads to weaker binding of HSA molecules to CB. Rate constants of DTNB reaction with the SH group of HSA applied to a CB column, bound-HSA and unbound-HSA fractions, were 4.8 x 10(-3), 21.6 x 10(-3), and 11.2 x 10(-3) s(-1), respectively. The HSA-SH group of diabetics is more reactive compared with control individuals (rate constants 20.9 x 10(-3)+/- 4.4 x 10(-3) vs 12.9 x 10(-3)+/- 2.6 x 10(-3) s(-1), P lt 0.05). Recovery values of the SH group obtained after chromatography of HSA with bound stearic acid ranged from 110 to 140%, while those for defatted HSA were from 98.5 to 101.7%. Thus, HSA-bound FA leads to an increase of HSA-SH content and a contribution to total serum thiols, which make the determination of the thiol group unreliable.
PB  - Academic Press Inc Elsevier Science, San Diego
T2  - Analytical Biochemistry
T1  - The influence of fatty acids on determination of human serum albumin thiol group
EP  - 57
SP  - 50
VL  - 448
DO  - 10.1016/j.ab.2013.11.030
UR  - conv_3171
ER  - 
@article{
author = "Jovanović, Vesna B. and Pavićević, Ivan D. and Takić, Marija M. and Penezic-Romanjuk, Ana Z. and Aćimović, Jelena M. and Mandić, Ljuba M.",
year = "2014",
abstract = "During investigation of the changes of the Cys34 thiol group of human serum albumin (HSA) (isolated by affinity chromatography with Cibacron Blue (CB)) in diabetes, we found that the HSA-SH content was higher (11-33%) than the total serum thiol content. The influence of fatty acids (FA) binding to HSA on this discrepancy was investigated in vitro (using fluorescence and CD spectroscopy and GC) and with HSA samples from diabetic (n=20) and control groups (n=17). HSA-bound FA determine the selection of HSA molecules by CB and enhance reactivity and/or accessibility of the SH group. A high content of polyunsaturated FA (35.6%) leads to weaker binding of HSA molecules to CB. Rate constants of DTNB reaction with the SH group of HSA applied to a CB column, bound-HSA and unbound-HSA fractions, were 4.8 x 10(-3), 21.6 x 10(-3), and 11.2 x 10(-3) s(-1), respectively. The HSA-SH group of diabetics is more reactive compared with control individuals (rate constants 20.9 x 10(-3)+/- 4.4 x 10(-3) vs 12.9 x 10(-3)+/- 2.6 x 10(-3) s(-1), P lt 0.05). Recovery values of the SH group obtained after chromatography of HSA with bound stearic acid ranged from 110 to 140%, while those for defatted HSA were from 98.5 to 101.7%. Thus, HSA-bound FA leads to an increase of HSA-SH content and a contribution to total serum thiols, which make the determination of the thiol group unreliable.",
publisher = "Academic Press Inc Elsevier Science, San Diego",
journal = "Analytical Biochemistry",
title = "The influence of fatty acids on determination of human serum albumin thiol group",
pages = "57-50",
volume = "448",
doi = "10.1016/j.ab.2013.11.030",
url = "conv_3171"
}
Jovanović, V. B., Pavićević, I. D., Takić, M. M., Penezic-Romanjuk, A. Z., Aćimović, J. M.,& Mandić, L. M.. (2014). The influence of fatty acids on determination of human serum albumin thiol group. in Analytical Biochemistry
Academic Press Inc Elsevier Science, San Diego., 448, 50-57.
https://doi.org/10.1016/j.ab.2013.11.030
conv_3171
Jovanović VB, Pavićević ID, Takić MM, Penezic-Romanjuk AZ, Aćimović JM, Mandić LM. The influence of fatty acids on determination of human serum albumin thiol group. in Analytical Biochemistry. 2014;448:50-57.
doi:10.1016/j.ab.2013.11.030
conv_3171 .
Jovanović, Vesna B., Pavićević, Ivan D., Takić, Marija M., Penezic-Romanjuk, Ana Z., Aćimović, Jelena M., Mandić, Ljuba M., "The influence of fatty acids on determination of human serum albumin thiol group" in Analytical Biochemistry, 448 (2014):50-57,
https://doi.org/10.1016/j.ab.2013.11.030 .,
conv_3171 .

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