Stable and Functional Rhomboid Proteases in Lipid Nanodiscs by Using Diisobutylene/Maleic Acid Copolymers

被引：36

作者：

Barniol-Xicota, Marta ^{[1
]}

Verhelst, Steven H. L. ^{[1
,2
]}

机构：

[1] KU Leuven Univ Leuven, Dept Cellular & Mol Med, Lab Chem Biol, Herestr 49 Box 802, B-3000 Leuven, Belgium

[2] AG Chem Prote, Leibniz Inst Analyt Sci ISAS, Otto Hahn Str 6b, D-44227 Dortmund, Germany

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2018年 / 140卷 / 44期

关键词：

MEMBRANE-PROTEINS; BILAYER NANODISCS; CRYSTAL-STRUCTURE; INTRAMEMBRANE; PROTEOLYSIS; FAMILY; REVEALS; ACTIVATION; INHIBITORS; DISCOVERY;

D O I：

10.1021/jacs.8b08441

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Rhomboid proteases form a paradigm for intramembrane proteolysis and have been implicated in several human diseases. However, their study is hampered by difficulties in solubilization and purification. We here report on the use of polymers composed of maleic acid and either diisobutylene or styrene for solubilization of rhomboid proteases in lipid nanodiscs, which proceeds with up to 48% efficiency. We show that the activity of rhomboids in lipid nanodiscs is closer to that in the native membrane than rhomboids in detergent. Moreover, a rhomboid that was proteolytically unstable in detergent turned out to be stable in lipid nanodiscs, underlining the benefit of using these polymer-stabilized nanodiscs. The systems are also compatible with the use of activity-based probes and can be used for small molecule inhibitor screening, allowing several downstream applications.

引用

页码：14557 / 14561

页数：5

共 33 条

[1] Mammalian EGF receptor activation by the rhomboid protease RHBDL2 [J].

Adrain, Colin ;

Strisovsky, Kvido ;

Zettl, Markus ;

Hu, Landian ;

Lemberg, Marius K. ;

Freeman, Matthew .

EMBO REPORTS, 2011, 12 (05) :421-427

[2] Two Plasmodium rhomboid proteases preferentially cleave different adhesins implicated in all invasive stages of malaria [J].

Baker, Rosanna P. ;

Wijetilaka, Ruvini ;

Urban, Sinisa .

PLOS PATHOGENS, 2006, 2 (10) :922-932

[3]

Baker RP, 2012, NAT CHEM BIOL, V8, P759, DOI [10.1038/nchembio.1021, 10.1038/NCHEMBIO.1021]

[4] Proteolysis inside the Membrane Is a Rate-Governed Reaction Not Driven by Substrate Affinity [J].

Dickey, Seth W. ;

Baker, Rosanna P. ;

Cho, Sangwoo ;

Urban, Sinisa .

CELL, 2013, 155 (06) :1270-1281

[5] Discovery and validation of 2-styryl substituted benzoxazin-4-ones as a novel scaffold for rhomboid protease inhibitors [J].

Goel, Parul ;

Jumpertz, Thorsten ;

Ticha, Anezka ;

Ogorek, Isabella ;

Mikles, David C. ;

Hubalek, Martin ;

Pietrzik, Claus U. ;

Strisovsky, Kvido ;

Schmidt, Boris ;

Weggen, Sascha .

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2018, 28 (08) :1417-1422

[6] Discovery and Biological Evaluation of Potent and Selective N-Methylene Saccharin-Derived Inhibitors for Rhomboid Intramembrane Proteases [J].

Goel, Parul ;

Jumpertz, Thorsten ;

Mikles, David C. ;

Ticha, Anezka ;

Nguyen, Minh T. N. ;

Verhelst, Steven ;

Hubalek, Martin ;

Johnson, Darren C. ;

Bachovchin, Daniel A. ;

Ogorek, Isabella ;

Pietrzik, Claus U. ;

Strisovsky, Kvido ;

Schmidt, Boris ;

Weggen, Sascha .

BIOCHEMISTRY, 2017, 56 (51) :6713-6725

[7] Membrane Proteins Solubilized Intact in Lipid Containing Nanoparticles Bounded by Styrene Maleic Acid Copolymer [J].

Knowles, Timothy J. ;

Finka, Rachael ;

Smith, Corinne ;

Lin, Yu-Pin ;

Dafforn, Tim ;

Overduin, Michael .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2009, 131 (22) :7484-+

[8] The rhomboids: a nearly ubiquitous family of intramembrane serine proteases that probably evolved by multiple ancient horizontal gene transfers [J].

Koonin, EV ;

Makarova, KS ;

Rogozin, IB ;

Davidovic, L ;

Letellier, MC ;

Pellegrini, L .

GENOME BIOLOGY, 2003, 4 (03)

[9] Regulated intracellular ligand transport and proteolysis control EGF signal activation in Drosophila [J].

Lee, JR ;

Urban, S ;

Garvey, CF ;

Freeman, M .

CELL, 2001, 107 (02) :161-171

[10] Encapsulated membrane proteins: A simplified system for molecular simulation [J].

Lee, Sarah C. ;

Khalid, Syma ;

Pollock, Naomi L. ;

Knowles, Tim J. ;

Edler, Karen ;

Rothnie, Alice J. ;

Thomas, Owen R. T. ;

Dafforn, Timothy R. .

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2016, 1858 (10) :2549-2557

← 1 2 3 4 →