Abstract
Hydrophobic interactions between nine model proteins and net-neutral lipid bilayer membranes (liposomes) under stress conditions were quantitatively examined by using immobilized liposome chromatography (ILC). Small or large unilamellar liposomes were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and immobilized in a gel matrix by utilizing covalent coupling between amino-containing lipids and activated gel beads or avidin-biotin biospecific binding. Retardation of bovine carbonic anhydrase (CAB) in ILC was pronounced at particular temperatures (50 and 60 °C) where the local hydrophobicity of theses protein molecules becomes sufficiently large. Protein-induced leakage of a hydrophilic dye (calcein) from immobilized liposomes interior was also drastically enhanced at particular temperatures where large retardation was observed. For other proteins examined, similar results were also observed. The specific capacity factor of the proteins characteristic for the ILC and the amount of calcein released from immobilized liposomes were successfully expressed as a function of the product of the local hydrophobicities of proteins and liposomes, regardless of protein species and the type of the stress conditions applied (denaturant and heating). These findings indicate that lipid membranes have an ability to non-specifically recognize local hydrophobicities of proteins to form stress-mediated supramolecular assemblies with proteins, which may have potential applications in bioprocesses such as protein refolding and separation. ILC was thus found to be a very useful method for the quantitative detection of dynamic protein-liposome interactions triggered by stress conditions.
| Original language | English |
|---|---|
| Pages (from-to) | 174-181 |
| Number of pages | 8 |
| Journal | Biochemical Engineering Journal |
| Volume | 29 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Apr 15 2006 |
| Externally published | Yes |
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Keywords
- Biosensors
- Bioseperations
- Immobilized liposome chromatography
- Membrane stress biotechnology
- Protein
- Protein denaturation
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Chemical Engineering(all)
Cite this
Evaluation of temperature and guanidine hydrochloride-induced protein-liposome interactions by using immobilized liposome chromatography. / Yoshimoto, Noriko; Yoshimoto, Makoto; Yasuhara, Kazuma; Shimanouchi, Toshinori; Umakoshi, Hiroshi; Kuboi, Ryoichi.
In: Biochemical Engineering Journal, Vol. 29, No. 3, 15.04.2006, p. 174-181.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Evaluation of temperature and guanidine hydrochloride-induced protein-liposome interactions by using immobilized liposome chromatography
AU - Yoshimoto, Noriko
AU - Yoshimoto, Makoto
AU - Yasuhara, Kazuma
AU - Shimanouchi, Toshinori
AU - Umakoshi, Hiroshi
AU - Kuboi, Ryoichi
PY - 2006/4/15
Y1 - 2006/4/15
N2 - Hydrophobic interactions between nine model proteins and net-neutral lipid bilayer membranes (liposomes) under stress conditions were quantitatively examined by using immobilized liposome chromatography (ILC). Small or large unilamellar liposomes were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and immobilized in a gel matrix by utilizing covalent coupling between amino-containing lipids and activated gel beads or avidin-biotin biospecific binding. Retardation of bovine carbonic anhydrase (CAB) in ILC was pronounced at particular temperatures (50 and 60 °C) where the local hydrophobicity of theses protein molecules becomes sufficiently large. Protein-induced leakage of a hydrophilic dye (calcein) from immobilized liposomes interior was also drastically enhanced at particular temperatures where large retardation was observed. For other proteins examined, similar results were also observed. The specific capacity factor of the proteins characteristic for the ILC and the amount of calcein released from immobilized liposomes were successfully expressed as a function of the product of the local hydrophobicities of proteins and liposomes, regardless of protein species and the type of the stress conditions applied (denaturant and heating). These findings indicate that lipid membranes have an ability to non-specifically recognize local hydrophobicities of proteins to form stress-mediated supramolecular assemblies with proteins, which may have potential applications in bioprocesses such as protein refolding and separation. ILC was thus found to be a very useful method for the quantitative detection of dynamic protein-liposome interactions triggered by stress conditions.
AB - Hydrophobic interactions between nine model proteins and net-neutral lipid bilayer membranes (liposomes) under stress conditions were quantitatively examined by using immobilized liposome chromatography (ILC). Small or large unilamellar liposomes were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and immobilized in a gel matrix by utilizing covalent coupling between amino-containing lipids and activated gel beads or avidin-biotin biospecific binding. Retardation of bovine carbonic anhydrase (CAB) in ILC was pronounced at particular temperatures (50 and 60 °C) where the local hydrophobicity of theses protein molecules becomes sufficiently large. Protein-induced leakage of a hydrophilic dye (calcein) from immobilized liposomes interior was also drastically enhanced at particular temperatures where large retardation was observed. For other proteins examined, similar results were also observed. The specific capacity factor of the proteins characteristic for the ILC and the amount of calcein released from immobilized liposomes were successfully expressed as a function of the product of the local hydrophobicities of proteins and liposomes, regardless of protein species and the type of the stress conditions applied (denaturant and heating). These findings indicate that lipid membranes have an ability to non-specifically recognize local hydrophobicities of proteins to form stress-mediated supramolecular assemblies with proteins, which may have potential applications in bioprocesses such as protein refolding and separation. ILC was thus found to be a very useful method for the quantitative detection of dynamic protein-liposome interactions triggered by stress conditions.
KW - Biosensors
KW - Bioseperations
KW - Immobilized liposome chromatography
KW - Membrane stress biotechnology
KW - Protein
KW - Protein denaturation
UR - http://www.scopus.com/inward/record.url?scp=33645992423&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33645992423&partnerID=8YFLogxK
U2 - 10.1016/j.bej.2005.08.030
DO - 10.1016/j.bej.2005.08.030
M3 - Article
AN - SCOPUS:33645992423
VL - 29
SP - 174
EP - 181
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
SN - 1369-703X
IS - 3
ER -