Variation of the detergent-binding capacity and phospholipid content of membrane proteins when purified in different detergents.

Ilgü, Hüseyin; Jeckelmann, Jean-Marc; Gachet Otanez, Maria Salomé; Boggavarapu, Venkata Ravi Naga Rajendr; Ucurum Fotiadis, Zöhre; Gertsch, Jürg; Fotiadis, Dimitrios José (2014). Variation of the detergent-binding capacity and phospholipid content of membrane proteins when purified in different detergents. Biophysical journal, 106(8), pp. 1660-1670. Biophysical Society 10.1016/j.bpj.2014.02.024

[img]
Preview
Text
Biophysical Journal Vol 106.pdf - Published Version
Available under License Publisher holds Copyright.

Download (2MB) | Preview

Purified membrane proteins are ternary complexes consisting of protein, lipid, and detergent. Information about the amounts of detergent and endogenous phospholipid molecules bound to purified membrane proteins is largely lacking. In this systematic study, three model membrane proteins of different oligomeric states were purified in nine different detergents at commonly used concentrations and characterized biochemically and biophysically. Detergent-binding capacities and phospholipid contents of the model proteins were determined and compared. The insights on ternary complexes obtained from the experimental results, when put into a general context, are summarized as follows. 1), The amount of detergent and 2) the amount of endogenous phospholipids bound to purified membrane proteins are dependent on the size of the hydrophobic lipid-accessible protein surface areas and the physicochemical properties of the detergents used. 3), The size of the detergent and lipid belt surrounding the hydrophobic lipid-accessible surface of purified membrane proteins can be tuned by the appropriate choice of detergent. 4), The detergents n-nonyl-β-D-glucopyranoside and Cymal-5 have exceptional delipidating effects on ternary complexes. 5), The types of endogenous phospholipids bound to membrane proteins can vary depending on the detergent used for solubilization and purification. 6), Furthermore, we demonstrate that size-exclusion chromatography can be a suitable method for estimating the molecular mass of ternary complexes. The findings presented suggest a strategy to control and tune the numbers of detergent and endogenous phospholipid molecules bound to membrane proteins. These two parameters are potentially important for the successul crystallization of membrane proteins for structure determination by crystallographic approaches.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Ilgü, Hüseyin; Jeckelmann, Jean-Marc; Gachet Otanez, Maria Salomé; Boggavarapu, Venkata Ravi Naga Rajendr; Ucurum Fotiadis, Zöhre; Gertsch, Jürg and Fotiadis, Dimitrios José

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

0006-3495

Publisher:

Biophysical Society

Language:

English

Submitter:

Barbara Järmann-Bangerter

Date Deposited:

02 Apr 2015 08:59

Last Modified:

30 Jan 2016 16:42

Publisher DOI:

10.1016/j.bpj.2014.02.024

PubMed ID:

24739165

BORIS DOI:

10.7892/boris.65713

URI:

https://boris.unibe.ch/id/eprint/65713

Actions (login required)

Edit item Edit item
Provide Feedback