Abstract
The transition of the holo-form of bovine α-lactalbumin from the native (N) to the pH-generated acidic-state (A-state) was analyzed by probing its tertiary and secondary structure using a concerted spectroscopic approach combining near- and far-UV circular dichroism (CD), electrospray ionization ion mobility mass spectrometry (ESI-IM-MS), vibrational circular dichroism (VCD), and Fourier transform infrared spectroscopy (FTIR) in the attenuated total reflection (ATR) and transmission (TR) modes. The spectroscopic results, which relied on the interaction of an electromagnetic field with different molecular targets, confirmed the decay of extensive rigid side-chain packing interactions during the pH-induced N → A-state transition and revealed the targets' dependence on secondary structural changes. Independent analyses of the spectral changes using two methods of multivariate analysis, such as principal component analysis and two-dimensional correlation spectroscopy, revealed small but significant differences in the secondary structure as a result of the all-or-none transition. The cooperativity of the transition was quantitatively described using values corresponding to the mid-point (tm) and width of the transition (Δtm). The averages of the two parameters, calculated using the data collected by the different probes, were equal to 3.5 ± 0.2 and 0.6 ± 0.1(SE), respectively. The variable two-state nature of the cooperative N → A-state transition confirmed that the protonation of the side chain carboxyl groups on the Asp and Glu residues and that the release of a Ca2 + ion induced structural changes on both the secondary and tertiary levels. The changes have been confirmed by results obtained from the concerted spectroscopic approach.
| Original language | English |
|---|---|
| Pages (from-to) | 593-606 |
| Number of pages | 14 |
| Journal | Biochimica et Biophysica Acta - Proteins and Proteomics |
| Volume | 1844 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2014 Mar 1 |
| Externally published | Yes |
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Keywords
- A-state molten globule
- Holo α-lactalbumin
- Multi-spectral analysis
- Principal component analysis
- Protein structure
- Two-dimensional correlation spectroscopy
ASJC Scopus subject areas
- Analytical Chemistry
- Biophysics
- Biochemistry
- Molecular Biology
Cite this
The transition from the native to the acid-state characterized by multi-spectroscopy approach : Study for the holo-form of bovine α-lactalbumin. / Litwińczuk, Adriana; Ryu, Soo Ryeon; Nafie, Laurence A.; Lee, Jong Wha; Kim, Hugh I.; Jung, Young Mee; Czarnik-Matusewicz, Bogusława.
In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1844, No. 3, 01.03.2014, p. 593-606.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The transition from the native to the acid-state characterized by multi-spectroscopy approach
T2 - Study for the holo-form of bovine α-lactalbumin
AU - Litwińczuk, Adriana
AU - Ryu, Soo Ryeon
AU - Nafie, Laurence A.
AU - Lee, Jong Wha
AU - Kim, Hugh I.
AU - Jung, Young Mee
AU - Czarnik-Matusewicz, Bogusława
PY - 2014/3/1
Y1 - 2014/3/1
N2 - The transition of the holo-form of bovine α-lactalbumin from the native (N) to the pH-generated acidic-state (A-state) was analyzed by probing its tertiary and secondary structure using a concerted spectroscopic approach combining near- and far-UV circular dichroism (CD), electrospray ionization ion mobility mass spectrometry (ESI-IM-MS), vibrational circular dichroism (VCD), and Fourier transform infrared spectroscopy (FTIR) in the attenuated total reflection (ATR) and transmission (TR) modes. The spectroscopic results, which relied on the interaction of an electromagnetic field with different molecular targets, confirmed the decay of extensive rigid side-chain packing interactions during the pH-induced N → A-state transition and revealed the targets' dependence on secondary structural changes. Independent analyses of the spectral changes using two methods of multivariate analysis, such as principal component analysis and two-dimensional correlation spectroscopy, revealed small but significant differences in the secondary structure as a result of the all-or-none transition. The cooperativity of the transition was quantitatively described using values corresponding to the mid-point (tm) and width of the transition (Δtm). The averages of the two parameters, calculated using the data collected by the different probes, were equal to 3.5 ± 0.2 and 0.6 ± 0.1(SE), respectively. The variable two-state nature of the cooperative N → A-state transition confirmed that the protonation of the side chain carboxyl groups on the Asp and Glu residues and that the release of a Ca2 + ion induced structural changes on both the secondary and tertiary levels. The changes have been confirmed by results obtained from the concerted spectroscopic approach.
AB - The transition of the holo-form of bovine α-lactalbumin from the native (N) to the pH-generated acidic-state (A-state) was analyzed by probing its tertiary and secondary structure using a concerted spectroscopic approach combining near- and far-UV circular dichroism (CD), electrospray ionization ion mobility mass spectrometry (ESI-IM-MS), vibrational circular dichroism (VCD), and Fourier transform infrared spectroscopy (FTIR) in the attenuated total reflection (ATR) and transmission (TR) modes. The spectroscopic results, which relied on the interaction of an electromagnetic field with different molecular targets, confirmed the decay of extensive rigid side-chain packing interactions during the pH-induced N → A-state transition and revealed the targets' dependence on secondary structural changes. Independent analyses of the spectral changes using two methods of multivariate analysis, such as principal component analysis and two-dimensional correlation spectroscopy, revealed small but significant differences in the secondary structure as a result of the all-or-none transition. The cooperativity of the transition was quantitatively described using values corresponding to the mid-point (tm) and width of the transition (Δtm). The averages of the two parameters, calculated using the data collected by the different probes, were equal to 3.5 ± 0.2 and 0.6 ± 0.1(SE), respectively. The variable two-state nature of the cooperative N → A-state transition confirmed that the protonation of the side chain carboxyl groups on the Asp and Glu residues and that the release of a Ca2 + ion induced structural changes on both the secondary and tertiary levels. The changes have been confirmed by results obtained from the concerted spectroscopic approach.
KW - A-state molten globule
KW - Holo α-lactalbumin
KW - Multi-spectral analysis
KW - Principal component analysis
KW - Protein structure
KW - Two-dimensional correlation spectroscopy
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UR - http://www.scopus.com/inward/citedby.url?scp=84893358278&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2013.12.018
DO - 10.1016/j.bbapap.2013.12.018
M3 - Article
C2 - 24389233
AN - SCOPUS:84893358278
VL - 1844
SP - 593
EP - 606
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 3
ER -