Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Santos, Hadassa Campos; Lamas, Regino Piñeiro; Ciancaglini, Pietro

doi:10.1590/s0100-879x2002000300002

Cited by 40 publications

(65 citation statements)

References 37 publications

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“…Many of the so-called integral or transmembrane proteins completely cross the lipidic bilayer and, due to the strong hydrophobic interaction with the phospholipids, can only be removed with the use of detergents-tensoactive compounds which have the ability to decrease the lipid-lipid and lipidprotein interactions during the protein solubilization process (Gennis 1989;Le Maire et al 2000;Santos and Ciancaglini 2000;Santos et al 2002;Magalhães et al 2003;Schuck et al 2003;Heerklotz 2008). On the other hand, peripheral or extrinsic proteins have a superficial interaction with the polar heads of the membrane lipids and can be removed simply by a change in ionic strength or pH.…”

Section: Purification or Expression Of The Membrane Proteinmentioning

confidence: 99%

“…These multilamellar systems can be sonicated or forced to go through (extrusion) pores of nanometric dimensions, to form unilamellar systems (with a single concentric bilayer), very similar to the natural membranes. The sonication as well as the extrusion process supplies the necessary energy to break the different lamellae and to re-organize the bilayers in uniform and control-sized vesicles (New 1990;Camolezi et al 1999Camolezi et al , 2002Ierardi et al 2002;Daghastanli et al 2004;Santos et al 2002Santos et al , 2005Rigos et al 2008;dos Santos et al 2010).…”

Section: Constructions Of Lipid Mimetic Systemsmentioning

confidence: 99%

“…It has been demonstrated that the αβ form of the Na,K-ATPase is capable of both ATP hydrolysis and active ion transport (Ward and Cavieres 1993, Martin et al 2000Miles et al 2011). However, accumulating evidence suggests that the enzyme normally self associates as (αβ) 2 dimers (Santos et al 2002(Santos et al , 2006bThoenges and Schoner 1997;Antolovic et al 1999;Costa et al 2003;Santos and Ciancaglini 2003) or as (αβ) 4 tetramers (Mahaney et al 1990;Donnet et al 2001;Taniguchi et al 2001).…”

Section: Applications For the Study Of Lipid-protein Interactionsmentioning

confidence: 99%

“…For example, good results for Na,K-ATPase reconstitution were obtained using a 1:1 (w/w) DPPC:DPPE mixture with a lipid:protein 1:3 (w/w) ratio resulting in proteoliposomes containing 89% of protein with average diameter of 140 nm and 79% recovered ATPase activity. Integrity studies of the Na,K-ATPase in this liposomes system have demonstrated that the enzyme is reconstituted with the ATP hydrolysis site located at the external side of the lipid bilayer vesicle, called inside-out orientation (Santos et al 2002(Santos et al , 2006b). These inside-out proteoliposomes enable a series of biophysical studies on the stability and structure of the enzyme.…”

Section: Applications For the Study Of Lipid-protein Interactionsmentioning

confidence: 99%

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Proteoliposomes in nanobiotechnology

et al. 2012

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Proteoliposomes are systems that mimic lipid membranes (liposomes) to which a protein has been incorporated or inserted. During the last decade, these systems have gained prominence as tools for biophysical studies on lipid-protein interactions as well as for their biotechnological applications. Proteoliposomes have a major advantage when compared with natural membrane systems, since they can be obtained with a smaller number of lipidic (and protein) components, facilitating the design and interpretation of certain experiments. However, they have the disadvantage of requiring methodological standardization for incorporation of each specific protein, and the need to verify that the reconstitution procedure has yielded the correct orientation of the protein in the proteoliposome system with recovery of its functional activity. In this review, we chose two proteins under study in our laboratory to exemplify the steps necessary for the standardization of the reconstitution of membrane proteins in liposome systems: (1) alkaline phosphatase, a protein with a glycosylphosphatidylinositol anchor, and (2) Na,K-ATPase, an integral membrane protein. In these examples, we focus on the production of the specific proteoliposomes, as well as on their biochemical and biophysical characterization, with emphasis on studies of lipid-protein interactions. We conclude the chapter by highlighting current prospects of this technology for biotechnological applications, including the construction of nanosensors and of a multiprotein nanovesicular biomimetic to study the processes of initiation of skeletal mineralization.

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Section: Purification or Expression Of The Membrane Proteinmentioning

confidence: 99%

Section: Constructions Of Lipid Mimetic Systemsmentioning

confidence: 99%

Section: Applications For the Study Of Lipid-protein Interactionsmentioning

confidence: 99%

Section: Applications For the Study Of Lipid-protein Interactionsmentioning

confidence: 99%

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Proteoliposomes in nanobiotechnology

et al. 2012

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“…For example, MLVs may be sonicated or passed through a membrane with pores of nanometric dimensions (extrusion method) to form unilamellar vesicles that have properties very similar those of to natural membranes. Both the sonication and extrusion methods provide the energy required to break the various lamellae that initially formed and reorganize the system into uniform bilayers with controlled sizes of vesicles (Camolezi et al 2002;Ierardi et al 2002;Daghastanli et al 2004;Santos et al 2002Santos et al , 2005Rigos et al 2008;dos Santos et al 2009;Bolean et al 2010Bolean et al , 2011Simão et al 2010a, b;Ciancaglini et al 2012).…”

Section: Liposomesmentioning

confidence: 99%

Liposomal systems as carriers for bioactive compounds

et al. 2015

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Since the revolutionary discovery that phospholipids can form closed bilayered structures in aqueous systems, the study of liposomes has become a very interesting area of research. The versatility and amazing biocompatibility of liposomes has resulted in their wide-spread use in many scientific fields, and many of their applications, especially in medicine, have yielded breakthroughs in recent decades. Specifically, their easy preparation and various structural aspects have given rise to broadly usable methodologies to internalize different compounds, with either lipophilic or hydrophilic properties. The study of compounds with potential biotechnological application(s) is generally related to evaluation and risk assessment of the possible cytotoxic or therapeutic effects of the compound under study. In most cases, undesirable sideeffects are associated with an interaction of the liposome with the cell membrane and/or its absorption and subsequent interaction with a cellular biomolecule. Liposomal carrier systems have an unprecedented potential for delivering bioactive substances to specific molecular targets due to their biocompatibility, biodegradability and low toxicity. Liposomes are therefore considered to be an invaluable asset in applied biotechnology studies due to their potential for interaction with both hydrophilic and lipophilic compounds.

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Biologically Inspired Hybrid Nanodevices

Patti

Montemagno

2007

Nanobiotechnology II

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The exploration and exploitation of biologic modes of design and self-assembly are now studies of both greater urgency and ease, as the tools for manipulation and visualization of nanoscale materials become increasingly available. Engineering biologically inspired nanoscale devices encompasses a wide variety of research, from current nanomaterials such as gecko tape, self-cleaning glass, and artificial shark skin [1], to the mechanics of how biological molecules such as proteins, enzymes, DNA and RNA can function as analogous man-made structures . In this chapter, we will briefly examine a sample of these technologies and follow up with current related research. We will discuss what can be gleaned from these emerging technologies, focusing primarily on biomimetic proteinbased devices. Finally, we will present our research efforts in the area of biocomputation and extend this discussion to the prospects of future applications.Engineering hybrid nanoscale devices requires the concurrent application of technology from a variety of fields. Incorporating varying levels of organization is central to creating functional biomimetic materials. The common thread among biological nano-hybrid devices is the need to exploit natural self-assembly schemes that have evolved over the millennia to build complex structures. Often, utilizing such a self-assembly scheme is sufficient, but optimum form may not always follow natural function, and producing devices which convert chemical energy to electricity, or light into chemical energy such as ATP can improve on Nature's design through engineering. Here, we show two bodies of work, including protein-based devices and cellular power generation, the common theme being a fusion of biologic molecules with synthetic structures to produce nanoscale hybrid devices. 401 Nanobiotechnology II. Edited by Chad A. Mirkin and Christof M. Niemeyer

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Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Cited by 40 publications

References 37 publications

Proteoliposomes in nanobiotechnology

Proteoliposomes in nanobiotechnology

Liposomal systems as carriers for bioactive compounds

Biologically Inspired Hybrid Nanodevices

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