The influence of fatty acids on determination of human serum albumin thiol group
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2014
Authors
Jovanović, Vesna B.
Pavićević, Ivan D.

Takić, Marija M.

Penezic-Romanjuk, Ana Z.

Aćimović, Jelena M.

Mandić, Ljuba M.

Article (Published version)

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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 / DiabetesSource:
Analytical Biochemistry, 2014, 448, 50-57Publisher:
- 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
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Institut za medicinska istraživanjaTY - 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 .