The human body is a complex mechanism and it basically is made up macro and micronutrients. The most important macronutrients are proteins, carbohydrates and some bit of fat. Micronutrients are also important though they are required in exceedingly small quantities. Vitamins and minerals form micronutrients. In this article we will try and understand more about protein kinase. We will find out what exactly they are and why they are considered important for our body. We are sure that it will help readers to have a better understanding of this chemical component and the important role it must play in our day to activities, growth and other such things.
What Is Protein Kinase?
Protein kinases are a special type of enzymes. Known as PTKs in its short form; they have many bodily functions. They help in regulating the various biological activities of protein intake that we go through daily. It helps in phosphorylation of certain types of amino acids. It makes use of ATP as the main source of phosphate. This helps many inactive forms of protein to turn into active forms. It can help in better synthesis of protein in the body and therefore it could ensure several benefits and advantages.
What Is Human Genome?
For better understanding of PTK we need to have some understanding about the human genome. We need to bear in mind that the human genome has around 500 odd protein kinase genes. They roughly contribute to around 2% of all the human genes in our body. It would also be pertinent to mention here that protein kinase is also found in plants and bacteria. The role of protein kinase is extremely important. This is because it helps to modify almost 30% of the total amount of human protein in the human body and this done through different types of kinase activities. They also have the capability of regulating the cellular pathways. They play a big role in regulating the cellular pathways of those activities that are related to signal transduction.
Why Is Protein Kinase Important?
The unique thing about protein kinase is that leads to a process called phosphorylation. We have talked about it briefly. However, there are a few things that make this process quite unique and different. It leads to something known as posttranslational modification. This can help to modulate protein role in many different physiological processes. This happens in almost in very foreseeable manner. It leads to minute and intrinsic activities at the biological level. Further it could also lead to changes at the sub-cellular location levels. It could also play a big role in determining the half-life when it comes to binding with other proteins. According to researchers and scientists, protein kinase is critical for regulating the function of key amino-acid components. It could help play a proactive as well as positive role as far as cell cycle progression is concerned.
What Are the Advantages of Phosphorylation?
When phosphorylation of a phosphate group happens to some polypeptide residues, it results in a covalent binding. This process of phosphorylation and dephosphorylation comes with several advantages and benefits. This is because it is extremely fast acting and rapid. Further the process can happen without new protein that may require degradation. Further the whole process is reversible and that is what dephosphorylation is all about. It also leads to structural changes that go a long way in regulating various protein functions. It does by better modulation of protein folding and removes affinity wherever need. Further, it also could add to the overall activity and stability.
There have been studies that go to prove that dephosphorylation is an efficient way of bringing about switch like changes in protein function. This also may lead to major changes as far as catalytic functioning of enzymes is concerned. These include phosphatases and kinases. It also often results in better rearrangement of the protein structure. It can bring about changes at the sub-cellular levels and improve interaction thereof.
It Could Help Regulate Various Functions
Phosphorylation from protein kinase also works exceedingly well as a molecular switch for various types of regulatory events. It helps to create the right signalling pathways. Further, it also helps to create the right balance between protein phosphorylation. In other words, it means that the cells can be able to compartmentalize both protein phosphatases and kinases. This process of separation or compartmentalization can also effectively and efficiently control the activities of various other protein and enzymes. It also could lead to the occurrence of various other posttranslational modifications.
Let us look at a simple example like budding yeast which is the one of the most primary forms of organism. It would be pertinent to mention here that around 3% of the proteins in yeast are phosphates or kinases. It would also not be out of place to mention that a few of these enzymes present in yeast are extremely specific. In other words, they have the capacity to phosphorylate or even dephosphorylate only a few specific target proteins. These target proteins include structural proteins, enzymes, ion channels, cell receptors, and signalling factors. When the protein activity is controlled while it is in a phosphorylation state, it will have certain obvious activities and changes. However, this activity would depend on the level and efficiency of kinases activity that work on it. The process is a continuous one as the phosphate group will be continuing to be added or removed. It is continuous and ongoing cycle that allows protein to switch rapidly and move from one state to another.
Why Is It So Much Talked About?
There are many reasons why protein kinase and the resultant changes are becoming so popular and so much talked about. These include reversibility, simplicity, and flexibility as far as phosphorylation is concerned. It is now being continuously adopted as one of the most trusted, proven and efficient control mechanisms at the cellular level. It has the capability of precisely regulating various events in space and time and as far as the entire process of cell progression is concerned.
What Are the Methods of Detecting Protein Phosphorylation Or Protein Kinase
Protein kinases have the capability of transferring phosphate from various groups. These include tyrosine, threonine, serine and various other protein peptide substrates. As mentioned above, almost around 30% of proteins that are present in the eukaryotic cells go through the process of phosphorylation. It has the capacity of regulating a wide variety of cellular activities. These include differentiation, cell cycle, metabolism and also highly critical neuronal communication.
However, it is important to identify and have some idea about the various methods that are being currently used as far as identifying this process is concerned.
KAA or Kinase Activity Assays
This is a method of assessing the activity of a specific and single kinase. It may be able to provide the right information as far as the parallel pathways are concerned. The biological sample to find out the exact kinase activity is measured by incubation. It is done by a process of immuno-precipitation of kinase. An exogenous substrate with ATP presence is used. However, this may not be the most accurate way of finding out more about the activity of single protein kinase.
PSAD or Phospho-Specific Antibody Development
This is an old and classic method that is used for direct measurement of protein phosphorylation. This process involves incubation of whole cells. This is done using 32P-orthophosphate that is radio labelled. This is a time consuming and labour-intensive method. The whole process would require hours of incubation and would require the use of different types of radioisotopes. There are other methods that also may be used. These include gel electrophoresis (2-dimensional). Some laboratories also use other types of phosphorylation techniques, but this could change the mobility as well as the isoelectric point as far as the protein is concerned.
Western Blot Test
This is a common and well-known method that is used for finding out more about the state of phosphorylation of protein. It is comparatively easier and the testing can be done over a widespread area. This is perhaps because most of the cell biology laboratories have the necessary equipment and infrastructure to perform these sophisticated experiments. The biological sample is transferred to a membrane for identifying the protein of interest. It would also be interesting to note that this form of test eliminates the problems and hazards of various disposal requirements.
Those who are in touch with modern day laboratory tests would understand that ELISA is the most reliable and powerful method for measuring protein kinase that leads to protein phosphorylation. This form of testing is quite reliable and more quantitative and qualitative when compared to Western Blotting and other forms of testing. The format is proven and it has been time-tested for years. It makes use of an antibody that is specific to the desired protein. It is also independent of the state of phosphorylation. The target protein must be purified or should be in a complex heterogeneous state and a common example is cell lysate.
Protein Kinase and its actions and manifestations are quite complex and complicated. Not many people belonging to the scientific and medical fraternity are fully aware of it. We hope and believe that the above article may have been able to give the readers a reasonably good insight into the various aspects of protein kinase from a layman’s perspective.
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