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    Cell biology

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
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    3 views4 pages

    Plasma Membrane

    Uploaded by

    hm5314581
    Cell biology

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
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    Spa BAPPADITYA PaTRA, m Se ( ation v) CNTOSENETICS £ MOLECULAR BloLomy BAPPADITYA PATRA, M.Se. ZOOLOGY M-7685965125 / 8902159352 - 200 BIO BASIC BAPPADITYA PATRA, M.Sc. ZOOLOGY BAPPADITYA PATRA, M.Sc. ZOOLOGY M-7685965125 / 8902159352 i ania CYTOLOGY : PLASMA MEMBRANE Robertson (1959) propounded his “Unit membrane hypothesis” in which he stated that all cellular membranes have a similar trilaminar structure consisting of a lipid bilayer sandwiched between two layers of proteins, Besides, based on the data available from x-ray diffraction studies, he further held that a membrane is about 7.5 nm thick, the lipid bilayer hhas a thickness of 35 nm and each pro- tein layer is 2 nm in thickness. Moreover, the proteins exist as f-pleated sheets over the hydrophilic heads of lipids. Robertson further proposed that the pro- teins on two surfaces of lipid bilayer may differ in nature. Thus, he first thought of asymmetric distribution of membrane proteins, Roberston’s model is practically a reiteration of Danielli-Davson model with extension of that model for all cellular membranes and with some additional idea on the organisation of membrane proteins. However, subsequent studies using optical rotatory dispersion, infrared spectroscopy and Circular dichroism revealed that membrane proteins do not exist as B-pleated sheets, but instead, they are mostly globular proteins having chelical organisation. (AZ Fuld mosaic model of Singer and QL Nicolson : The concept of molecular > organisation of cell membranes as pro- pounded by Danielli and Davson in the 1930s and Robertson in the 1950s to 1960s underwent significant reorganisation fol- lowing the accumulation of data from enzymatic studies, freeze-fracture stu- dies, ESR (Electron Spin) spectroscopic studies and different other types. of sophisticated studies on membranes, Based on such data, $. Jonathan Singer and Garth Nicolson of the University of California (1972) propounded their ‘Fluid mosacic model’ for membrane organisation. This model had served as the ‘central dogma’ of membrane biolo- gy for more than two decades. The basic postulates of this model are as follows (ig. 32): U0) (The membrane is not a rigid, static structure but a quasi-fluid structure) is of a fluid consistency like oil.(The cellular mem- brane is a dynamic structure in which the components are mobile and capable of com- ing together to engage in various types of transient interactions, The membrane is not just a sandwich of lipid bilayer within two. layers of protein, but instead isa mossie rangement of discontinuous protein molecules within lipid bilayer. On account of its fluidity and the mosaic arrangement of protein molecules, this model of membrane Structure is Known as the ‘fluid mosaic model’, and it describes both properties as well as organisation of the membrane. U2) The lipid molecules of the lipid bilayer have their hydrophilic or polar heads facing outward and the hydrophobic tails facing the membrane interior (Fig. 6) Membrane proteins are of two types. Integral proteins remain embedded in the lipid bilayer upto various depths and peri- pheral proteins.remain bound to the outer or Inner surface of the lipid bilayer. Some of the integral proteins are transmembrane proteins passing across the lipid bilayer. The peri- pheral’ proteins remain associated with membranes by ionic interaction with the hydrophilic heads of the lipid bilayer. The integral proteins have two parts — the hydrophilic part remains close to the polar head groups of lipid, while the hydrophobic part remains buried in the lipid bilayer due to hydrophobic interaction with the hydro- phobic tails of lipids. @) Membrane proteins are mostly globu- lar proteins having c-helical organisation instead of being B-pleated sheets, \6) Both the lipid and integral protein molecules are capable of free movements within the plane of the membrane, This model has received support from various experimental and analytical studies, Evidences in favour of Fluid mosaic model A. Direct evidence from enzymatic studies on mosaic arrangement Enzymatic studies by Singer and Nicol- son have clearly shown that the proteins do not form continuous layers over the lipid bilayer and instead, may be embedded upto various depths into the lipid bilayer. Isolated plasma membranes were subjec- ted to hydrolytic action of phospholipase which hydrolyses phospholipids. This enzyme was chosen since the phospholipids constitute the main bulk of the total lipid fraction of most membranes. Nearly 60-70% of total mem- brane lipids were removed from the mem- branes and passed into the medium. The pro- tein fraction of membranes remained unatfec- ted and could not be detected in the medium. That a major fraction of the membrane lipids was extracted by the hydrolytic action of the enzyme confirms that the lipid bilayer was not completely covered by protein layers, In otherwards, the protein molecules remain embedded in the lipid bilayer instead of forming continuous sheaths on the surfaces of the lipid bilayer. B. Freeze fracture studies of membranes Direct evidence in favour of the fluid mosaic model comes from freeze fracture studies of the membrane. This technique not only reveals that the lipid bilayer is not sand- wiched between two continuous layers of proteins, but also reveals the asymmetric dis- tribution of membrane proteins as well as mobility of membrane proteins. Under EM, a replica produced from freeze-fractured RBC ghosts revealed that numerous protrusions are randomly spread on the replica. The protrusions represent the locations of membrane proteins that remained embedded in the lipid bilayer. The protru- sions were more numerous in the inner or protoplasmic half or P-face of the membrane than in the external half or E-face of the membrane. This study clearly indicates that protein particles retain embedded in the lipid bilayer of membranes and are asymme- trically distributed in it (Fig. 3.3). Evidences on fluidity Different experimental studies con- firmed the mobility of both membrane lipids and membrane proteins ah ae exterior half ne st APRN 35 eADITE LOS 21 carbohydrate pay costs L thain\ wee? ? proteins : ey Geirecure plasma { es membrane |! protein; ' 1 i ' teytoplasm \ v\ E --\) protoplasmic half Fig. 3.3: Diagrammatic representati the centre of the lipid bilay. ion of a freeze fractured plasma membrane. The fracture plane (arrow) occurs at ver and passes over (or under) the integral membrane proteins. 200 BIO BASIC BAPPADITYA PATRA, M.Sc. ZOOLOGY BAPPADITYA PATRA, M.Sc. ZOOLOGY 768.59 5 8902159352 eyTOLocy : Pas Ma biz tiReae25 / #90215 AA. Fluidity or mobility of membrane the fluorescent dye fluorescein which gives a lipids : Studies based on biophysical techniques like “ESR Spectroscopy’ and ‘Differential scanning colorimetry” revealed that mem- brane lipids are capable of two kinds of movements (i) lateral movements (diffusion) vithin the respective monolayers and Mi) flip-flop movement or transbilayer move- ment where lipids move across the bilayer. Lateral movements of lipid molecule are quite fast than the flip-flop movement (Fig. 3.4). Individual lipid molecules can rotate very rapidly around their own axis and their hydrocarbon chains show flexion. feral diffusion f j », mae ab) IN } occurs) rotation - flexion Fig. 3.4: ‘The types of movement possible for phospho- lipid molecules in a lipid bilayer, In low temperature, the lipid molecules become immobile, since their hydrocarbon chains remain in a ‘gel state’ or rigid ‘crys- talline state’. But at physiological tempera- ture, the lipid molecules become mobile due to rotaion of carbon-carbon bonds caused by heat energy. 6. Fluidity or movements of membrane proteins Various experimental studies have confirmed that’ the (membrane proteins are actually capable of lateral movements or Jateral diffusions within the lipid bilayer) “Cell f 1970) : In this experiment, particular membrane proteins of mouse fibroblast cells were labelled with specific antibodie ion” experiment (Frye and E linked with green fluorescence. Particular membrane proteins of human fibroblast cells were labelled with specific antibodies linked with the fluorescent dye, rhodamine, which gives a red fluorescence. The labelled mouse cells and human cells were then induced to fuse together in vitro by treatment with Sendai virus, The resultant fused cells or hybrid cells were kept under incubation at 37°C and af} * {o>} ys

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