Running head: EFFECT OF MEMBRANE INTERACTIONS ON RAS PROTEIN ACTIVITY

Effect of Membrane Interactions on Ras Protein Activity

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Abstract

The Ras proteins play a crucial role in regulating key cellular processes such as differentiation,

apoptosis, and proliferation. Most of the localization of the Ras occurs in the plasma membrane

in which most of the interactions take place. A tight regulation of the spatiotemporal dynamics of

Ras proteins is, for example, due to the vital oncogenic mediators informing the specific activity

of the proteins. Ras undergoes lipid raft association processes and membrane localization both of

which draw from the different membrane interactions. An examination of the spatiotemporal Ras

organization within the cell membrane is evident in extensive studies. The membrane

interactions might be having an impact on protein signaling activity of Ras. Nanocluster

formation depends of membrane interactions and forms a crucial part of Ras signaling.

Introduction

Ras proteins represent the GTPases thought to oscillate between the different bound

states and the play role of molecular switches in signaling pathways (Zhou & Hancock, 2015).

This work seeks to understand the role of the membrane linkers in the activity of Ras proteins

and the possible effect of Ras protein entry into cholesterol rich regions on the activity of

specific Ras proteins. The proteins have a low molecular mass implicated in mediation of

signaling processes originating from the receptors of the cell surface (Eisenberg et al., 2011).

While Ras proteins occur in a localized state within the plasma membranes, the proteins also

occur in other organelles such as the mitochondria and endoplasmic reticulum (Lin et al., 2014).

The plasma membrane represents the primary signaling platform for Ras in which the cell

membrane interactions influence the activity of the different isoforms of Ras during signal

transduction.
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Effect of Membrane Interactions

The N-Ras protein develops active and preferential interaction with most of the

cholesterol-dependent components and assemblies using in vivo observations. The N-Ras

interactions and the resulting dynamics within the cell membranes of live cells helps in

understanding the dependence on the activated state of the plasma membrane. Eisenberg et al.

(2011) determined that the interactions of N-Ras relies on activation of the plasma membrane by

expressing the protein in cells following a beam size analysis using the fluorescence recovery

after photobleaching (FRAP) approach. The investigation demonstrates an instance in which N-

Ras- GTP depalmitoylation stimulation dependents on the cross-linking of raft proteins

(Eisenberg et al., 2013). The dissociation of Ras from the plasma membrane raft clusters relies

on the cross-linking process leading to protein accumulation in and Golgi compartment

signaling.

The differential interactions between plasma membrane’s raft clusters and the Ras

proteins informs the level of specificity evident in the effects of raft clustering. The GTP loaded

form attracts a preferential targeting from N-Ras proteins. The mechanisms linking selective

interactions and raft clustering allow an instance of co-stimulation due to the cross-linked raft

proteins (Sarkar-Banerjee et al., 2017). The interactions play a role in altering the spatial and

kinetic patterns expressed in the Ras protein in terms of signaling and activation following

primary stimulation. For instance, Zhou and Hancock (2017) suggest that the long-term signaling

reduces from the plasma membrane following a kinetic and spatial activation of the Ras protein.

The interactions between Ras and cell membranes together with signaling undergoes the

modulation initiated by cross-linking and raft clustering.

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A lipid-anchored contributes to the signaling node within cells. Observations involving

Ras and the influence of membrane interactions its activity indicate that the organization of Ras

occurs within the cell membrane, contributing to the regulatory role (Lin, et al., 2014). The Ras

protein organization and interactions within the cell membrane draws from the activities of K-

Ras, and the moieties of farnesyl and palmitoyl. For instance, observations suggest the formation

of a dimer between H-Ras and the cell membrane surface resulting in an interface of protein-

protein binding (Lin et al., 2014). Findings indicate H-Ras influences rotational and transitional

mobility within supported membrane. The protein attaches to the cell membrane through a

coupling process involving various cysteines known as C184 and C181 in the hypervariable

region of the maleimide-functionalized lipid.

The rotational and translational mobility of the Ras protein particularly the H-Ras

dependents of the surface and density. The measurements made using the fluorescence

correlation spectroscopy (FCS) concerning the lateral diffusion rates of the protein as a function

of the cell membrane surface density indicate the effect on protein mobility (Lin et al., 2014).

The increase in the surface density correlates with a similar increase on the lateral transition

mobility of the Ras proteins. The rate of diffusion of the proteins across the cell membrane in

terms of the rotational mobility appears to increase significantly with an increase in the cell

membrane surface density. The lipid-anchored Ras protein experiences unrestricted lateral

diffusion mobility with the domination of cell membrane component properties using in vitro

and in vivo investigations (Zhou at al., 2017). The interactions leading to high membrane

clustering of the Ras protein results in lower mobility. Apart from increased protein clustering,

additional protein-lipid interactions lead to significant reductions in mobility of the protein.

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Zhou and Hancock (2015) examined the interactions and demonstrated the existence of a

complex spatiotemporal Ras distribution within the plasma membrane. The fluorescence imaging

approaches reveal a significantly dynamic protein organization and subsequent interactions with

the cell membrane. With over 60% of the proteins diffusing in the form of free mobile

monomers, the remaining proportion has immobile nanodomians. The interactions involving the

formation of dimers play a critical role in larger nanoclusters assembly process.

Studies focusing on the lipid composition of Ras helps in understanding the effect of their

entry within the different cholesterol-rich regions. Some experiments on the stability of

nanoclusters of Ras in conditions of depleted cholesterol suggest that cholesterol dependent

nanoclusters form with the H-Ras GDP. On the contrary, The H-Ras GTP undergoes a targeted

segregation to form nanoclusters with cholesterol independent characteristics. The findings

underscore the importance of cholesterol distribution within specific forms of the nanoclusters

formed from the Ras protein. For instance, Zhou and Hancock (2015) showed that signaling

transduction and binding of Ras relies on the incorporation of cholesterol with clusters of Ras.

Vogel et al. (2014) reported similar findings on N-Ras protein interaction with the raft model

membranes with varying extends of complexity. The domain boundaries experience increased

accumulation of N-Ras due to the resulting line tension produced by Ras protein entry into

cholesterol rich regions.

Conclusion

A complex relationship exists between the plasma membrane and the Ras protein forms.

The different isoforms of Ras such as the N-Ras, K-Ras, and H-Ras interact with the plasma

membrane through the formation of nanoclusters, dimers, and spatial organization. An active
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preferential interaction with cholesterol is demonstrated in both in vitro and in vivo

investigations. In particular, the H-Ras GDP isoform develops cholesterol dependent

nanoclusters based on experiments involving cholesterol depletion. It is quite interesting to note

that not all forms of Ras are cholesterol dependent. For instance, the H-Ras GTP form has

cholesterol independent features and undergoes observable segregation during the formation of

its nanoclusters. The analysis of the influence of membrane interactions on the Ras protein

activity depicts an instance the rotational and translational mobility depending on parameters

such as the surface density. When the plasma membrane surface density increases, the rate of

Ras protein diffusion rises. A lateral and unrestricted diffusion occurs for all lipid-anchored Ras.

The future work may seek to establish the precise mechanism linking selective membrane

interaction and raft clustering with the different isoforms of Ras protein.
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Bibliography

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