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Platelet glycoprotein VI

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The cytosol , also known as cytoplasmic matrix or groundplasm , is one of the liquids found inside cells ( intracellular fluid (ICF)). It is separated into compartments by membranes. For example, the mitochondrial matrix separates the mitochondrion into many compartments.

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58-447: 2GI7 51206 243816 ENSG00000278316 ENSG00000275633 ENSG00000274566 ENSG00000275931 ENSG00000276065 ENSMUSG00000078810 Q9HCN6 P0C191 NM_016363 NM_001083899 NM_001256017 NM_001163014 NP_001077368 NP_001242946 NP_057447 NP_001156486 Platelet glycoprotein VI ( GPVI ) is a glycoprotein receptor for collagen which

116-501: A molecular mass of less than 300  Da . This mixture of small molecules is extraordinarily complex, as the variety of molecules that are involved in metabolism (the metabolites ) is immense. For example, up to 200,000 different small molecules might be made in plants, although not all these will be present in the same species, or in a single cell. Estimates of the number of metabolites in single cells such as E. coli and baker's yeast predict that under 1,000 are made. Most of

174-416: A high viscosity , for example, in egg white and blood plasma . Variable surface glycoproteins allow the sleeping sickness Trypanosoma parasite to escape the immune response of the host. The viral spike of the human immunodeficiency virus is heavily glycosylated. Approximately half the mass of the spike is glycosylation and the glycans act to limit antibody recognition as the glycans are assembled by

232-409: A large central cavity that is isolated from the remainder of the cytosol. One example of such an enclosed compartment is the proteasome . Here, a set of subunits form a hollow barrel containing proteases that degrade cytosolic proteins. Since these would be damaging if they mixed freely with the remainder of the cytosol, the barrel is capped by a set of regulatory proteins that recognize proteins with

290-585: A low concentration of sodium ions. This difference in ion concentrations is critical for osmoregulation , since if the ion levels were the same inside a cell as outside, water would enter constantly by osmosis - since the levels of macromolecules inside cells are higher than their levels outside. Instead, sodium ions are expelled and potassium ions taken up by the Na⁺/K⁺-ATPase , potassium ions then flow down their concentration gradient through potassium-selection ion channels, this loss of positive charge creates

348-421: A much denser meshwork of actin fibres than the remainder of the cytosol. These microdomains could influence the distribution of large structures such as ribosomes and organelles within the cytosol by excluding them from some areas and concentrating them in others. The cytosol is the site of multiple cell processes. Examples of these processes include signal transduction from the cell membrane to sites within

406-537: A negative membrane potential . To balance this potential difference , negative chloride ions also exit the cell, through selective chloride channels. The loss of sodium and chloride ions compensates for the osmotic effect of the higher concentration of organic molecules inside the cell. Cells can deal with even larger osmotic changes by accumulating osmoprotectants such as betaines or trehalose in their cytosol. Some of these molecules can allow cells to survive being completely dried out and allow an organism to enter

464-416: A set of proteins with similar functions, such as enzymes that carry out several steps in the same metabolic pathway. This organization can allow substrate channeling , which is when the product of one enzyme is passed directly to the next enzyme in a pathway without being released into solution. Channeling can make a pathway more rapid and efficient than it would be if the enzymes were randomly distributed in

522-677: A signal directing them for degradation (a ubiquitin tag) and feed them into the proteolytic cavity. Another large class of protein compartments are bacterial microcompartments , which are made of a protein shell that encapsulates various enzymes. These compartments are typically about 100–200 nanometres across and made of interlocking proteins. A well-understood example is the carboxysome , which contains enzymes involved in carbon fixation such as RuBisCO . Non-membrane bound organelles can form as biomolecular condensates , which arise by clustering, oligomerisation , or polymerisation of macromolecules to drive colloidal phase separation of

580-462: A state of suspended animation called cryptobiosis . In this state the cytosol and osmoprotectants become a glass-like solid that helps stabilize proteins and cell membranes from the damaging effects of desiccation. The low concentration of calcium in the cytosol allows calcium ions to function as a second messenger in calcium signaling . Here, a signal such as a hormone or an action potential opens calcium channel so that calcium floods into

638-725: Is a compound containing carbohydrate (or glycan) covalently linked to protein. The carbohydrate may be in the form of a monosaccharide, disaccharide(s). oligosaccharide(s), polysaccharide(s), or their derivatives (e.g. sulfo- or phospho-substituted). One, a few, or many carbohydrate units may be present. Proteoglycans are a subclass of glycoproteins in which the carbohydrate units are polysaccharides that contain amino sugars. Such polysaccharides are also known as glycosaminoglycans. A variety of methods used in detection, purification, and structural analysis of glycoproteins are The glycosylation of proteins has an array of different applications from influencing cell to cell communication to changing

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696-424: Is attached to the protein in a cotranslational or posttranslational modification . This process is known as glycosylation . Secreted extracellular proteins are often glycosylated. In proteins that have segments extending extracellularly, the extracellular segments are also often glycosylated. Glycoproteins are also often important integral membrane proteins , where they play a role in cell–cell interactions. It

754-418: Is complete, the amino acid sequence can be expanded upon using solid-phase peptide synthesis. Cytosol In the eukaryotic cell , the cytosol is surrounded by the cell membrane and is part of the cytoplasm , which also comprises the mitochondria, plastids , and other organelles (but not their internal fluids and structures); the cell nucleus is separate. The cytosol is thus a liquid matrix around

812-472: Is expressed in platelets . In humans, glycoprotein VI is encoded by the GP6 gene . It was first cloned in 2000 by several groups including that of Martine Jandrot-Perrus from INSERM. GPVI is a 58-kD platelet membrane glycoprotein that plays a crucial role in the collagen-induced activation and aggregation of platelets. Upon injury to the vessel wall and subsequent damage to the endothelial lining, exposure of

870-466: Is extremely high, and approaches 200 mg/ml, occupying about 20–30% of the volume of the cytosol. However, measuring precisely how much protein is dissolved in cytosol in intact cells is difficult, since some proteins appear to be weakly associated with membranes or organelles in whole cells and are released into solution upon cell lysis . Indeed, in experiments where the plasma membrane of cells were carefully disrupted using saponin , without damaging

928-789: Is important to distinguish endoplasmic reticulum-based glycosylation of the secretory system from reversible cytosolic-nuclear glycosylation. Glycoproteins of the cytosol and nucleus can be modified through the reversible addition of a single GlcNAc residue that is considered reciprocal to phosphorylation and the functions of these are likely to be an additional regulatory mechanism that controls phosphorylation-based signalling. In contrast, classical secretory glycosylation can be structurally essential. For example, inhibition of asparagine-linked, i.e. N-linked, glycosylation can prevent proper glycoprotein folding and full inhibition can be toxic to an individual cell. In contrast, perturbation of glycan processing (enzymatic removal/addition of carbohydrate residues to

986-530: Is likely to have been secondary to its role in host-pathogen interactions. A famous example of this latter effect is the ABO blood group system . Though there are different types of glycoproteins, the most common are N -linked and O -linked glycoproteins. These two types of glycoproteins are distinguished by structural differences that give them their names. Glycoproteins vary greatly in composition, making many different compounds such as antibodies or hormones. Due to

1044-508: Is not well understood. The concentrations of ions such as sodium and potassium in the cytosol are different to those in the extracellular fluid ; these differences in ion levels are important in processes such as osmoregulation , cell signaling , and the generation of action potentials in excitable cells such as endocrine, nerve and muscle cells. The cytosol also contains large amounts of macromolecules , which can alter how molecules behave, through macromolecular crowding . Although it

1102-451: Is that about 5% of this water is strongly bound in by solutes or macromolecules as water of solvation , while the majority has the same structure as pure water. This water of solvation is not active in osmosis and may have different solvent properties, so that some dissolved molecules are excluded, while others become concentrated. However, others argue that the effects of the high concentrations of macromolecules in cells extend throughout

1160-450: Is through the reaction between a protected glycan and a protected Asparagine. Similarly, an O-linked glycoprotein can be formed through the addition of a glycosyl donor with a protected Serine or Threonine . These two methods are examples of natural linkage. However, there are also methods of unnatural linkages. Some methods include ligation and a reaction between a serine-derived sulfamidate and thiohexoses in water. Once this linkage

1218-510: Is when the effective concentration of other macromolecules is increased, since they have less volume to move in. This crowding effect can produce large changes in both the rates and the position of chemical equilibrium of reactions in the cytosol. It is particularly important in its ability to alter dissociation constants by favoring the association of macromolecules, such as when multiple proteins come together to form protein complexes , or when DNA-binding proteins bind to their targets in

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1276-422: The genome . Although the components of the cytosol are not separated into regions by cell membranes, these components do not always mix randomly and several levels of organization can localize specific molecules to defined sites within the cytosol. Although small molecules diffuse rapidly in the cytosol, concentration gradients can still be produced within this compartment. A well-studied example of these are

1334-553: The "calcium sparks" that are produced for a short period in the region around an open calcium channel . These are about 2  micrometres in diameter and last for only a few milliseconds , although several sparks can merge to form larger gradients, called "calcium waves". Concentration gradients of other small molecules, such as oxygen and adenosine triphosphate may be produced in cells around clusters of mitochondria , although these are less well understood. Proteins can associate to form protein complexes , these often contain

1392-542: The body is mucins , which are secreted in the mucus of the respiratory and digestive tracts. The sugars when attached to mucins give them considerable water-holding capacity and also make them resistant to proteolysis by digestive enzymes. Glycoproteins are important for white blood cell recognition. Examples of glycoproteins in the immune system are: H antigen of the ABO blood compatibility antigens. Other examples of glycoproteins include: Soluble glycoproteins often show

1450-483: The cell, causing a decrease in drug effectiveness. Therefore, being able to inhibit this behavior would decrease P-glycoprotein interference in drug delivery, making this an important topic in drug discovery. For example, P-Glycoprotein causes a decrease in anti-cancer drug accumulation within tumor cells, limiting the effectiveness of chemotherapies used to treat cancer. Hormones that are glycoproteins include: Quoting from recommendations for IUPAC: A glycoprotein

1508-418: The cell, such as the cell nucleus , or organelles. This compartment is also the site of many of the processes of cytokinesis , after the breakdown of the nuclear membrane in mitosis . Another major function of cytosol is to transport metabolites from their site of production to where they are used. This is relatively simple for water-soluble molecules, such as amino acids, which can diffuse rapidly through

1566-443: The cytoplasm or nucleus. Although the cytoskeleton is not part of the cytosol, the presence of this network of filaments restricts the diffusion of large particles in the cell. For example, in several studies tracer particles larger than about 25  nanometres (about the size of a ribosome ) were excluded from parts of the cytosol around the edges of the cell and next to the nucleus. These "excluding compartments" may contain

1624-451: The cytoplasm that is contained within organelles. Due to the possibility of confusion between the use of the word "cytosol" to refer to both extracts of cells and the soluble part of the cytoplasm in intact cells, the phrase "aqueous cytoplasm" has been used to describe the liquid contents of the cytoplasm of living cells. Prior to this, other terms, including hyaloplasm , were used for the cell fluid, not always synonymously, as its nature

1682-410: The cytoskeleton by motor proteins . The cytosol is the site of most metabolism in prokaryotes, and a large proportion of the metabolism of eukaryotes. For instance, in mammals about half of the proteins in the cell are localized to the cytosol. The most complete data are available in yeast, where metabolic reconstructions indicate that the majority of both metabolic processes and metabolites occur in

1740-504: The cytosol and that water in cells behaves very differently from the water in dilute solutions. These ideas include the proposal that cells contain zones of low and high-density water, which could have widespread effects on the structures and functions of the other parts of the cell. However, the use of advanced nuclear magnetic resonance methods to directly measure the mobility of water in living cells contradicts this idea, as it suggests that 85% of cell water acts like that pure water, while

1798-566: The cytosol contains the cell's genome , within a structure known as a nucleoid . This is an irregular mass of DNA and associated proteins that control the transcription and replication of the bacterial chromosome and plasmids . In eukaryotes the genome is held within the cell nucleus , which is separated from the cytosol by nuclear pores that block the free diffusion of any molecule larger than about 10  nanometres in diameter. This high concentration of macromolecules in cytosol causes an effect called macromolecular crowding , which

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1856-433: The cytosol is water , which makes up about 70% of the total volume of a typical cell. The pH of the intracellular fluid is 7.4. while human cytosolic pH ranges between 7.0 and 7.4, and is usually higher if a cell is growing. The viscosity of cytoplasm is roughly the same as pure water, although diffusion of small molecules through this liquid is about fourfold slower than in pure water, due mostly to collisions with

1914-412: The cytosol, and can also prevent the release of unstable reaction intermediates. Although a wide variety of metabolic pathways involve enzymes that are tightly bound to each other, others may involve more loosely associated complexes that are very difficult to study outside the cell. Consequently, the importance of these complexes for metabolism in general remains unclear. Some protein complexes contain

1972-407: The cytosol. However, hydrophobic molecules, such as fatty acids or sterols , can be transported through the cytosol by specific binding proteins, which shuttle these molecules between cell membranes. Molecules taken into the cell by endocytosis or on their way to be secreted can also be transported through the cytosol inside vesicles , which are small spheres of lipids that are moved along

2030-407: The cytosol. This sudden increase in cytosolic calcium activates other signalling molecules, such as calmodulin and protein kinase C . Other ions such as chloride and potassium may also have signaling functions in the cytosol, but these are not well understood. Protein molecules that do not bind to cell membranes or the cytoskeleton are dissolved in the cytosol. The amount of protein in cells

2088-420: The glycan), which occurs in both the endoplasmic reticulum and Golgi apparatus , is dispensable for isolated cells (as evidenced by survival with glycosides inhibitors) but can lead to human disease (congenital disorders of glycosylation) and can be lethal in animal models. It is therefore likely that the fine processing of glycans is important for endogenous functionality, such as cell trafficking, but that this

2146-535: The host cell and so are largely 'self'. Over time, some patients can evolve antibodies to recognise the HIV glycans and almost all so-called 'broadly neutralising antibodies (bnAbs) recognise some glycans. This is possible mainly because the unusually high density of glycans hinders normal glycan maturation and they are therefore trapped in the premature, high-mannose, state. This provides a window for immune recognition. In addition, as these glycans are much less variable than

2204-399: The large numbers of macromolecules in the cytosol. Studies in the brine shrimp have examined how water affects cell functions; these saw that a 20% reduction in the amount of water in a cell inhibits metabolism, with metabolism decreasing progressively as the cell dries out and all metabolic activity halting when the water level reaches 70% below normal. Although water is vital for life,

2262-469: The most common cell line used for recombinant glycoprotein production is the Chinese hamster ovary line. However, as technologies develop, the most promising cell lines for recombinant glycoprotein production are human cell lines. The formation of the link between the glycan and the protein is key element of the synthesis of glycoproteins. The most common method of glycosylation of N-linked glycoproteins

2320-706: The most common. Monosaccharides commonly found in eukaryotic glycoproteins include: The sugar group(s) can assist in protein folding , improve proteins' stability and are involved in cell signalling. The critical structural element of all glycoproteins is having oligosaccharides bonded covalently to a protein. There are 10 common monosaccharides in mammalian glycans including: glucose (Glc), fucose (Fuc), xylose (Xyl), mannose (Man), galactose (Gal), N- acetylglucosamine (GlcNAc), glucuronic acid (GlcA), iduronic acid (IdoA), N-acetylgalactosamine (GalNAc), sialic acid , and 5- N-acetylneuraminic acid (Neu5Ac). These glycans link themselves to specific areas of

2378-453: The organelles. In prokaryotes , most of the chemical reactions of metabolism take place in the cytosol, while a few take place in membranes or in the periplasmic space . In eukaryotes, while many metabolic pathways still occur in the cytosol, others take place within organelles. The cytosol is a complex mixture of substances dissolved in water. Although water forms the large majority of the cytosol, its structure and properties within cells

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2436-407: The other cell membranes, only about one quarter of cell protein was released. These cells were also able to synthesize proteins if given ATP and amino acids, implying that many of the enzymes in cytosol are bound to the cytoskeleton. However, the idea that the majority of the proteins in cells are tightly bound in a network called the microtrabecular lattice is now seen as unlikely. In prokaryotes

2494-471: The production of the protein. Glycosylation is a process that roughly half of all human proteins undergo and heavily influences the properties and functions of the protein. Within the cell, glycosylation occurs in the endoplasmic reticulum . There are several techniques for the assembly of glycoproteins. One technique utilizes recombination . The first consideration for this method is the choice of host, as there are many different factors that can influence

2552-488: The protein amino acid chain. The two most common linkages in glycoproteins are N -linked and O -linked glycoproteins. An N -linked glycoprotein has glycan bonds to the nitrogen containing an asparagine amino acid within the protein sequence. An O -linked glycoprotein has the sugar is bonded to an oxygen atom of a serine or threonine amino acid in the protein. Glycoprotein size and composition can vary largely, with carbohydrate composition ranges from 1% to 70% of

2610-487: The purposes of this field of study is to determine which proteins are glycosylated and where in the amino acid sequence the glycosylation occurs. Historically, mass spectrometry has been used to identify the structure of glycoproteins and characterize the carbohydrate chains attached. The unique interaction between the oligosaccharide chains have different applications. First, it aids in quality control by identifying misfolded proteins. The oligosaccharide chains also change

2668-421: The remainder is less mobile and probably bound to macromolecules. The concentrations of the other ions in cytosol are quite different from those in extracellular fluid and the cytosol also contains much higher amounts of charged macromolecules such as proteins and nucleic acids than the outside of the cell structure. In contrast to extracellular fluid, cytosol has a high concentration of potassium ions and

2726-637: The resulting platelet plug is the initial hemostatic event limiting blood loss. However, exposure of collagen after rupture of atherosclerotic plaques is a major stimulus of thrombus formation associated with myocardial infarction or stroke. Complete or partial deficiency of GPVI in humans is a rare condition presenting as a mild bleeding disorder. GPVI has been shown to interact with LYN . Glycoprotein Glycoproteins are proteins which contain oligosaccharide (sugar) chains covalently attached to amino acid side-chains. The carbohydrate

2784-405: The solubility and polarity of the proteins that they are bonded to. For example, if the oligosaccharide chains are negatively charged, with enough density around the protein, they can repulse proteolytic enzymes away from the bonded protein. The diversity in interactions lends itself to different types of glycoproteins with different structures and functions. One example of glycoproteins found in

2842-449: The structure of this water in the cytosol is not well understood, mostly because methods such as nuclear magnetic resonance spectroscopy only give information on the average structure of water, and cannot measure local variations at the microscopic scale. Even the structure of pure water is poorly understood, due to the ability of water to form structures such as water clusters through hydrogen bonds . The classic view of water in cells

2900-507: The subendothelial matrix to blood flow results in deposition of platelets. Collagen fibers are the most thrombogenic macromolecular components of the extracellular matrix, with collagen types I, III, and VI being the major forms found in blood vessels. Platelet interaction with collagen occurs as a 2-step procedure: (1) the initial adhesion to collagen is followed by (2) an activation step leading to platelet secretion, recruitment of additional platelets, and aggregation. In physiologic conditions,

2958-493: The success of glycoprotein recombination such as cost, the host environment, the efficacy of the process, and other considerations. Some examples of host cells include E. coli, yeast, plant cells, insect cells, and mammalian cells. Of these options, mammalian cells are the most common because their use does not face the same challenges that other host cells do such as different glycan structures, shorter half life, and potential unwanted immune responses in humans. Of mammalian cells,

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3016-403: The thermal stability and the folding of proteins. Due to the unique abilities of glycoproteins, they can be used in many therapies. By understanding glycoproteins and their synthesis, they can be made to treat cancer, Crohn's Disease , high cholesterol, and more. The process of glycosylation (binding a carbohydrate to a protein) is a post-translational modification , meaning it happens after

3074-543: The total mass of the glycoprotein. Within the cell, they appear in the blood, the extracellular matrix , or on the outer surface of the plasma membrane, and make up a large portion of the proteins secreted by eukaryotic cells. They are very broad in their applications and can function as a variety of chemicals from antibodies to hormones. Glycomics is the study of the carbohydrate components of cells. Though not exclusive to glycoproteins, it can reveal more information about different glycoproteins and their structure. One of

3132-402: The underlying protein, they have emerged as promising targets for vaccine design. P-glycoproteins are critical for antitumor research due to its ability block the effects of antitumor drugs. P-glycoprotein, or multidrug transporter (MDR1), is a type of ABC transporter that transports compounds out of cells. This transportation of compounds out of cells includes drugs made to be delivered to

3190-400: The wide array of functions within the body, interest in glycoprotein synthesis for medical use has increased. There are now several methods to synthesize glycoproteins, including recombination and glycosylation of proteins. Glycosylation is also known to occur on nucleo cytoplasmic proteins in the form of O -GlcNAc . There are several types of glycosylation, although the first two are

3248-454: Was first introduced in 1965 by H. A. Lardy, and initially referred to the liquid that was produced by breaking cells apart and pelleting all the insoluble components by ultracentrifugation . Such a soluble cell extract is not identical to the soluble part of the cell cytoplasm and is usually called a cytoplasmic fraction. The term cytosol is now used to refer to the liquid phase of the cytoplasm in an intact cell. This excludes any part of

3306-427: Was not well understood (see protoplasm ). The proportion of cell volume that is cytosol varies: for example while this compartment forms the bulk of cell structure in bacteria , in plant cells the main compartment is the large central vacuole . The cytosol consists mostly of water, dissolved ions, small molecules, and large water-soluble molecules (such as proteins). The majority of these non-protein molecules have

3364-401: Was once thought to be a simple solution of molecules, the cytosol has multiple levels of organization. These include concentration gradients of small molecules such as calcium , large complexes of enzymes that act together and take part in metabolic pathways , and protein complexes such as proteasomes and carboxysomes that enclose and separate parts of the cytosol. The term "cytosol"

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