Heat Shock Proteins Lab Report

Heat shock proteins (HSPs), also known as stress inducing proteins exist in every organism inside of their cells. These proteins take part in very important roles such as, controlling cell cycle, cell signalling, protection against cell apoptosis, moving and sorting proteins into correct compartments, folding and unfolding proteins structures and building multi protein complexes. Therefore, they are very highly protected since they play all these key roles in cells and proteins (Li Z and Srivastava.P, 2004). This lab is concentrated on a specific heat shock protein called heat shock protein 70 (Hsp 70).
Hsp70s plays a crucial role in protein folding processes. The Hsp70s help assist the folding of proteins in three ways; stopping the clumping

Other JDP can stop the clumping of proteins on their own by quickly combining with the substrates or through ATP-independent transient. Hsp70 folding usually happens within minutes, and in this time interval the substrate cycle goes between free and bound stages constantly until the original state is achieved. During this substrate cycle there are two other options for proteins folding in the original state, which are keeping substrate concentration on the lower end to reduce clumping so the free molecules can achieve the original state by protein folding. As well as, during the constant free and bound stages causes the substrate to reverse its folding, for example, in the alpha and beta sheets, and then surpass the obstacles to be able to fold to the original state. In smaller amounts of clumped proteins can be completely solubilized by DnaK system itself. Although in larger amounts of clumping proteins the DnaK system only works a small amount, but when the Hsp70 binds itself to bigger clumping proteins the higher temperature dependence of mobility in the Hsp70 compared to the larger clumped protein.

All the different levels of gene regulation of the Hsp70 gene is discovered to be a reaction to heat stress, which could be due to increase in temperature, lower pH, protein unfolding or hypoxia. When heat stress stops the Hsp gene expression, then the Hsp70 mRNA expression begins (Silver and Noble, 2012).

The main techniques used in this experiment are; SDS-PAGE protein electrophoresis, coomassie staining and western blotting. SDS-PAGE is a regularly used procedure to separate proteins via electrophoresis using sodium dodecyl sulphate (SDS) to break down proteins, and a medium of discontinuous polyacrylamide gel. SDS-PAGE can be used for many different uses such as; finding important proteins in a sample and figuring out how much each protein is in a sample (Experimental Biosciences).

Coomassie staining is used on SDS-PAGE gels after proteins get separated by SDS-PAGE machine, in which the SDS-PAGE gels get immersed in coomassie stain and the extra amount gets rinsed off with solvent. When the staining of SDS PAGE is completed is should show blue bands on the clear background of the SDS PAGE gel (BIO RAD).
Western blotting is a lab procedure that is used to find specific proteins in a mixture