Amino Acid Hydrolysis

in this video, we're going to talk about our first protein cleavage technique Amino acid, hydraulics, ISS. So recall that way back in our previous lessons. When we talked about peptide bonds, we said that hydraulic sis is just a reaction in which bonds are cleaved with the treatment of water. And so amino acid hydraulics is is specifically when peptide bonds air cleaved with the treatment of water. And so, in our example below, we're gonna look at the hydraulics ISS of a peptide bond and which will notice is on the far left. Here. What we have is a dive peptide or ah, peptide with just two amino acid residues. And these two amino acid residues are linked via a peptide bond which is shown in pink right here. And so we know that if we treat our die peptide with water were able to induce hydrologists. But we also know from our previous lesson videos that hydraulics is is a very slow reaction. And so, in order to speed up the reaction, we can actually add an acid catalyst that will speed up the reaction. And so h plus here is representing our acid catalyst and so What you'll see is that our die peptide upon hydraulics is ends up forming to free amino acids, one free amino acid here on the far left and another free amino acid here on the right and so complete amino acid hydraulics. ISS can be induced with six Mohler hydrochloric acid and six Mohler hydrochloric acid essentially is going to non. Specifically, it will non specifically cleave all of the peptide bonds where all here is incredibly important and emphasized. And so what that results in is all of the constituent amino acids of the protein, uh, being released as free amino acids and so down below, in our example again, What we can see is that we have a protein here on the far left and notice that our protein has a bunch of amino acids which are represented by these circles. And we have an amino group, a free amino group on the N terminal end of our protein and a free car box, a group on the C terminal end of our protein. And when we treat our protein with six Mohler hydrochloric acid, we know that that is able to induce a complete amino acid hydraulics ISS, where all of the peptide bonds are going to be cleaved with the presence of water. So we know that most of our proteins are sitting in a solution and acquia solution that has water in it. And so, essentially, what will happen is all the peptide bonds are gonna be cleaved, and that releases all of the constituent free amino acids and so notice that they're no longer linked together via peptide bonds. Now there is a chemical known as men hydrant that will actually react with these free amino acids to produce a color for quantification. And so that allows us to quantify the number of amino acids that we have via light. Absorb INTs using a spectrum FA, Tom Attar and essentially, if we look down below, what you'll see is that we can treat our free amino acids over here with this chemical that's shown that's known as Nen Hydrant. And so in in hydrant will essentially react with all of these free amino acids to produce this bluish purplish color. And that bluish purplish color can be quantified with light, absorb INTs using a spectra fa Tom better, and that allows us to quantify the number of amino acids that we have. And then essentially, what we could do is we can take all of these free amino acids here, and we can analyze them with a technique such as mass spectrometry or UH H P L C in order to determine the protein composition or the amino acid composition. And so, in our example down below, which will notices were treating are or were analyzing? These amino acids here with H P, L C and H P L C essentially can allow us to reveal the amino acid composition of our protein, and so you can see that all of the amino acids have now been revealed through the use of H P L. C. Now what's really important, actually, is that once we use six Mohler hydrochloric acid, we're actually not able to determine the sequence of our protein. And so even though we are able to determine composition, which is the number and types of amino acids, we have no way to determine the exact order. So remember the sequence is referring to the exact order of amino acids from the N terminal end to the C terminal, and so what that means is, once we use six Mohler hydrochloric acid on a particular sample, we automatically lose the ability to determine the sequence of the residues. But we're able to pretty easily identify the composition, and so that's something that's important to keep in mind. So this here concludes our lesson on amino acid hydrologist, and we'll be able to get some practice utilizing thes concepts in our practice video, so I'll see you guys there.

What is the function of ninhydrin?

• A. Cleavage of proteins into free amino acids.
• B. Colorimetric agent to detect amino acids.
• C. Agent to carboxymethylate cysteines.
• D. Serves as the mobile phase in HPLC.