Calculating Isoelectric Point of Proteins (Example)

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  • @aarondesalvio
    @aarondesalvio4 жыл бұрын

    Thank you so much, this was explained incredibly well! One piece of advice for anyone who's confused: keep in mind whether or not the side chain is neutral or positive in the protonated form. This is relevant for the COO- group, where it is NEUTRAL when protonated, as opposed to an NH3 group that is POSITIVE when protonated.

  • @SouthpawC

    @SouthpawC

    3 жыл бұрын

    thanks!

  • @JuanRomero-xe2lf
    @JuanRomero-xe2lf8 жыл бұрын

    KZread Channels like this one-- (among a few others) are the future of learning. Thank you.

  • @absenzz
    @absenzz8 жыл бұрын

    Not all heroes wear capes...

  • @AKLECTURES

    @AKLECTURES

    8 жыл бұрын

    +Fab Gon Thanks bud! :)

  • @superoxidedismutase5757

    @superoxidedismutase5757

    6 жыл бұрын

    some wear AK lectures t-shirts

  • @charmainemoya2524
    @charmainemoya25248 жыл бұрын

    Wow, you're a LIFE SAVER. While I spent hours on homework trying to read my book and notes and still scratching my head, you make everything clear in less than 15 minutes per lesson. Love all your videos!

  • @lazer1235
    @lazer12359 жыл бұрын

    Very well explained, best video I have come across on youtube that explains this principle thus far

  • @AKLECTURES

    @AKLECTURES

    9 жыл бұрын

    lazer1235 Thanks! I appreciate that :)

  • @elenakhachaturyan8146

    @elenakhachaturyan8146

    8 жыл бұрын

    +lazer1235 Totally agree!!!

  • @chillstep4life
    @chillstep4life7 жыл бұрын

    Amazing! Very easy to understand, very well organized! Cant say enough about how clear and concise your lecture is. Thanks for the video.

  • @tylerlynn2655
    @tylerlynn26552 жыл бұрын

    this guy is the only reason I made it to pharmacy school

  • @Horsin4years
    @Horsin4years8 жыл бұрын

    This was super helpful. I like that you explain the steps very clearly and make a point to separate them out.

  • @andrewcho9779
    @andrewcho97798 жыл бұрын

    Thank you so much! I got very confused on this section and my professor wasn't of much help on figuring out how to to this but looking at this, I realize that it is much more simple than we think!

  • @cambokid16
    @cambokid168 жыл бұрын

    At first your videos were intimidating at first glance by the excessiveness, but omg it's very simple, straight to the point, and the explanation is beautiful, thank you!

  • @thepowerofpositivity8075
    @thepowerofpositivity80758 жыл бұрын

    Jus follow the simple rule if (1) the compound is a weak acid then the protonated form will be uncharged (AH) and the deprotonated form (A) will be negatively charged;(2) the compound is a weak base then the protonated form (BH) will be positively charged and the deprotonated form (B) will be uncharged. The tendency a compound will donate or accept proton depends on pH of the solution. Hope this help!

  • @funnysjoker
    @funnysjoker7 жыл бұрын

    Wow great lecture! Thanks so much! Keep the videos coming! I watched this on the bus, and it was very easy to follow. Thank you!

  • @thubelihlesithole5184
    @thubelihlesithole51848 жыл бұрын

    I'm a bit confused, I'm not quite sure why you concluded the two acidic amino acids as neutral and not positive.

  • @official.rajarshidutta

    @official.rajarshidutta

    3 жыл бұрын

    exactly my doubt

  • @kritankunwar4776

    @kritankunwar4776

    3 жыл бұрын

    @@official.rajarshidutta I think its because glutamate and aspartate was negatively charged hence oxygen has a negative charge to start with so when you add hydrogen that makes it neutral.

  • @pfeliciano4062

    @pfeliciano4062

    2 жыл бұрын

    So if the PH is within 1 unit away from the Pka the charge remains neutral. You see that we chose 3.5 as a guess for the PH, and the difference between that Ph=3.5 and Pka-4.1 is 0.6 and this is within the range of 1 unit away. You cannot have greater or less than that 1 unit. also if you are wondering why we did not just use a PH of 4.1, that is because for our second step we need to pick the pka values that are above and below our PH estimate, so as we knew we needed to pick a value within 1PH unit away from that Pka, we chose the PH close close enough to 4.1. Hope this helps and God bless your studying!

  • @sonjak8265

    @sonjak8265

    2 жыл бұрын

    @@pfeliciano4062 thank you

  • @yazan4mjad

    @yazan4mjad

    2 жыл бұрын

    Simply when pH goes lower than pKa of the carboxylic group which is 4.1, the carboxylic (-COO-) group will be protonated (-COOH) and it will have no charge, thus it is considered neutral.

  • @tanyaqualls9206
    @tanyaqualls92068 жыл бұрын

    Perfect! Thank you so much --this really helped me before my biochem exam! :)

  • @joseibarra9808
    @joseibarra98087 жыл бұрын

    Your videos are so helpful. Great for the new MCAT.

  • @britforlife32
    @britforlife328 жыл бұрын

    I didn't understand how to calculate pI before this, but now I do! Thank you so much, it's much appreciated

  • @penelopetaylor8384
    @penelopetaylor83847 жыл бұрын

    You are a true hero. THANK YOU for these amazing lectures.

  • @kissesVlogs17
    @kissesVlogs177 жыл бұрын

    After watching this, I might just pass my biochem exam. Thank you so much 😭😊

  • @crystalguerreromorris7102
    @crystalguerreromorris71025 жыл бұрын

    Your explanation was flawless, I finally comprehend!!!

  • @MultiSamSami
    @MultiSamSami6 жыл бұрын

    Man... I was looking for this. Thank you so much.

  • @pj10red
    @pj10red6 жыл бұрын

    Thank you soo much! My first Biochem exam is tomorrow and you saved me!!

  • @jacksonwestaway3376
    @jacksonwestaway33768 жыл бұрын

    Your videos are fantastic! Rlly appreciate them and you; getting me through MBLG =)

  • @simrankaur11
    @simrankaur118 жыл бұрын

    That was very well explained! Thank you.

  • @TheToxicMegacolon
    @TheToxicMegacolon4 жыл бұрын

    AK saving the day again for the 6th semester lol!

  • @amerain1729
    @amerain17296 жыл бұрын

    This is very helpful. Thank you!

  • @YaacovPsuturi
    @YaacovPsuturi4 ай бұрын

    I love u man, your explanation is perfect!!!

  • @jalehnikfarjam9844
    @jalehnikfarjam98443 жыл бұрын

    Thank you for the clear explanation!

  • @jacobboeckelman3691
    @jacobboeckelman36917 жыл бұрын

    This helped out so much. Thanks!

  • @gorkemnailaydn1524
    @gorkemnailaydn15242 жыл бұрын

    Dude you are awesome. Thank you so much for this understandable and good lesson.

  • @caiquebarreto6498
    @caiquebarreto64985 жыл бұрын

    Thank you for this awesome explaination!!!

  • @pratibhamali3845
    @pratibhamali38458 жыл бұрын

    this is very helpful to me.thank you so much.

  • @remyakin7198
    @remyakin71987 жыл бұрын

    You just saved a life!! Thanks

  • @buluchka
    @buluchka7 жыл бұрын

    I feel this will save my biochem grade, Thank you.

  • @syyylvo
    @syyylvo3 жыл бұрын

    Very clear and coincise!

  • @enricotortelli3642
    @enricotortelli36424 жыл бұрын

    You literally saved my life!

  • @shivansh709
    @shivansh7093 жыл бұрын

    Thanks bro! Helped me in my aits!

  • @zena1212
    @zena12128 жыл бұрын

    I finally get it thank you so much!

  • @reubent5780
    @reubent57809 жыл бұрын

    AK LECTURES Hey great video ..one thing that i'm a bit stuck on is why pka of 4.1 was neutral at pH 3.5 for the first example and why it was neutral for pka of 8.3 at ph7 in the 2nd example. thanks

  • @marchizaaa

    @marchizaaa

    9 жыл бұрын

    reuben Thomas george At pH=3.5, the Carboxylic group (pKa 4.1 in the picture) is neutral because is protonated,so you have a COOH group without charge. At pH=7 is the same...the SH group of the cysteine (pKa 8.3 in the picture) is neutral because it can do only 2 bonds, like an hydroxyl group ( OH), and because of this is neutral (it's difficult that O and S do 3 bonds,usually on organic molecules they are unstable in this state)...it has no charge. If we were at pH= 10 , the cysteine would be S- with a negative charge ( the -SH group would lose a proton because of OH anion forming H2O). I hope it is clear now. See ya!

  • @thepowerofpositivity8075

    @thepowerofpositivity8075

    8 жыл бұрын

    +reuben Thomas george Jus follow the simple rule if (1) the compound is a weak acid then the protonated form will be uncharged (AH) and the deprotonated form (A) will be negatively charged;(2) the compound is a weak base then the protonated form (BH) will be positively charged and the deprotonated form (B) will be uncharged. The tendency a compound will donate or accept proton depends on pH of the solution. Hope this help!

  • @the.angeltaylor

    @the.angeltaylor

    5 жыл бұрын

    @@marchizaaa I don't really see where the COOH group is pronated

  • @marchizaaa

    @marchizaaa

    5 жыл бұрын

    @@the.angeltaylor a COO- group (pKa = 4.1) gets protonation at pH 3.5. Therefore becomes COOH group, a neutral charged one.

  • @the.angeltaylor

    @the.angeltaylor

    5 жыл бұрын

    @@marchizaaa thankyou so much!!

  • @DefenderX
    @DefenderX5 жыл бұрын

    Is the pka value for each functional group in each amino acid accurate? Or do they vary depending on the peptide sequence and potential folding?

  • @AP-lj5qe
    @AP-lj5qe9 жыл бұрын

    amazing video, very clear explanation. I am glad to watch this video because my professor explanation is sucked.

  • @omarrosario1489
    @omarrosario14898 жыл бұрын

    Sir, why I did not find you earlier? Excellent video, may God protect you wherever you are and gives you health and happiness. Good vibes.

  • @SoraKimchi
    @SoraKimchi Жыл бұрын

    Thank u so much man❤

  • @yaarithanan1994
    @yaarithanan19946 жыл бұрын

    Thank you very much!

  • @zay7968
    @zay79688 жыл бұрын

    Thank you so much! I have my final exam tomorrow and this helped a lot! Hope u succeed in whatever you do.

  • @AKLECTURES

    @AKLECTURES

    8 жыл бұрын

    +Shadow walker Thanks! I appreciate that! best of luck on your exam, let me know how it all turns out!

  • @abayady1
    @abayady17 жыл бұрын

    This is a great video, thank you! However, I have a question, why don't you count the charges of amino acids individually? For example, Aspartate and Glutamate are both negatively charged aa, but you do not address their charge, and start as if they were neutral? My professor does count them. Would love to hear your feedback

  • @blehm8400

    @blehm8400

    6 жыл бұрын

    Basically, the easiest way I found to think about it is this. At a very low pH (e.g. pH 1) there is a high concentration of H+ ions (this is what pH measures). Therefore, anything that can accept a H+ ion will do so. This includes aspartate and glutamate which you'd normally think of as being negatively charged due to the COO- group they both have at the end of their side chains. When we raise the pH to 4.1 (the pKa value of aspartate and glutamate) they will accept a H+ ion and the COO- group will become COOH. So basically, start from a low pH , where everything that can be protonated (contains a H+ ion) is protonated. Look at the overall charge of the peptide. Then increase the pH and each group with a pKa value will be deprotonated (lose a H+ ion) at its pKa value. At each pKa value, note the new overall charge of the peptide. Keep doing this until you find the 2 pKa values that you will average to get the pI (isoelecric point). Hope that clairifes it.

  • @chamandeepkaur2177
    @chamandeepkaur21777 жыл бұрын

    Thank you so much. You're the best :-)

  • @ahalyalenka7358
    @ahalyalenka73588 жыл бұрын

    wow......... Lheninger ate my head....take my Namaskaar...

  • @chanchalmaji1456
    @chanchalmaji14567 жыл бұрын

    Again thanks for this

  • @AndreasChristodoulou99
    @AndreasChristodoulou993 жыл бұрын

    I love you for this video sir

  • @monday2471
    @monday24718 жыл бұрын

    do you exclude the pKas of the terminal groups when calculating the pI?

  • @Kka-ong-the-cat
    @Kka-ong-the-cat8 жыл бұрын

    Super helpful

  • @salam495
    @salam4958 жыл бұрын

    thank you alot !

  • @roblingbling
    @roblingbling7 жыл бұрын

    great videos!. If you figure out that a 15kDa protein has a PI of 9.71, and then it forms a homodimer. Does that affect the PI?

  • @catherinepaciotti3800
    @catherinepaciotti38006 жыл бұрын

    Hi, thanks for the video. Did you forget to take into account the pka of cysteine?

  • @asie5651
    @asie56518 жыл бұрын

    this guy had me thinking he's left handed for one minute 29 seconds.

  • @husamidrees4609
    @husamidrees46098 жыл бұрын

    Great video.

  • @niloofarfam4294
    @niloofarfam42947 жыл бұрын

    Thank you!!!!!

  • @tomasaguilar3017
    @tomasaguilar30176 жыл бұрын

    You make this too easy to understand it makes me question if Biochemistry is even a difficult subject......

  • @jadzgburella2068
    @jadzgburella20687 жыл бұрын

    Thanks ak lecture!!

  • @jadzgburella2068

    @jadzgburella2068

    7 жыл бұрын

    Getting my yoni all steamed up

  • @_Maya_Andrea
    @_Maya_Andrea7 жыл бұрын

    THANK YOU SO MUCH OH MY GOD

  • @walaahussein462
    @walaahussein4624 жыл бұрын

    Thanks for the video. How to calculate PI for modified peptides such as lantibiotics?

  • @lorcanokane5128
    @lorcanokane51283 жыл бұрын

    Much better than my professor

  • @hasanciftci9432
    @hasanciftci94324 жыл бұрын

    Amazing.

  • @crackers0413
    @crackers04138 ай бұрын

    Hi, I’m a little confused. The pka of the NH3 of Cysteine is 10.7, so averaging 4.07 of glutamic acid and 8.37 of cysteine gives 6.22

  • @junczhang
    @junczhang8 жыл бұрын

    thanks!!!

  • @joancalatayudvernich1237
    @joancalatayudvernich12379 жыл бұрын

    GREAT!

  • @RandomNooby
    @RandomNooby4 жыл бұрын

    @AK Re COVID Why is the death rate in Italy in 0-18 year olds less than 2% with everyone else roughly evenly divided into 3 equal groups. With many young adults to 50 year olds suffering from a cytokine storm, what are your thoughts?

  • @soumyamajumdar7929
    @soumyamajumdar79294 ай бұрын

    can you please find the pI for ala-arg-gly.

  • @ansammkh1681
    @ansammkh16812 жыл бұрын

    thank you

  • @TheMachomaniac
    @TheMachomaniac2 жыл бұрын

    Dont guess. Start with a fully protonated protein and solve towards a higher pH until you find the answer, a neutral peptide. The you find the pKa range and can solve the pI.

  • @the.angeltaylor
    @the.angeltaylor5 жыл бұрын

    Why would the charges for the first example be neutral?

  • @celestial4646
    @celestial46466 жыл бұрын

    thanks from a local ub student ;)

  • @ElizabethRey97
    @ElizabethRey979 жыл бұрын

    Thanks! I didn't even start college and I perfectly understood this :)

  • @joy397
    @joy3974 жыл бұрын

    thank you!!!!!

  • @shaboatrad4066
    @shaboatrad40668 жыл бұрын

    Aw.. why not using the pka of S-H which was 8.3 instead of 8.0 ?! Isn't that wrong?

  • @kareemm8608
    @kareemm86083 жыл бұрын

    My issue with a pH of 3 is that there will be a fractional charge in the COO- terminal so it won't necessarily be -1.

  • @sharmaeaselearningbiologyc5277
    @sharmaeaselearningbiologyc52774 жыл бұрын

    Sir thanku from india

  • @davehileri7072
    @davehileri70722 жыл бұрын

    Great👍

  • @gabrielwinces6606
    @gabrielwinces66066 жыл бұрын

    well explained

  • @tatendawarima4607
    @tatendawarima46079 жыл бұрын

    superb, thanks a lot!!!!!

  • @AKLECTURES

    @AKLECTURES

    9 жыл бұрын

    Tatenda Warima you're welcome :)

  • @dblkarno
    @dblkarno8 жыл бұрын

    how do you know that the pKa of the H2N of the Asp. is 8.0?

  • @lixx1414

    @lixx1414

    6 жыл бұрын

    My teacher gives them to us

  • @the.angeltaylor
    @the.angeltaylor5 жыл бұрын

    This is confusing, why do the rules apply to one part of the protein and not the others? What makes the Cysteine side chain group different that the others? What makes the pka of 4.1 in the first problem different than the other parts of the protein? Can somebody break this down please?

  • @davehileri7072
    @davehileri70722 жыл бұрын

    Ty

  • @rahimshah5859
    @rahimshah58596 жыл бұрын

    very helpfull

  • @mauimanman5498
    @mauimanman54985 жыл бұрын

    In the second problem where the pka is 8.3 why is it neutral?? wasn't it suppose to be positive because the pka was above the ph?

  • @MsTommyknocker
    @MsTommyknocker8 жыл бұрын

    I tried calculating the isoelectric point of the polypeptide with the following sequence: KRHKKDE. I got the pI to 11.51, but expasy protein parameter calculator got it to 9.74 or something like that. Could you clarify???

  • @jdevola999

    @jdevola999

    8 жыл бұрын

    +MrTommyknocker I agree -- it seems like none of these calculators agree. Why is that?

  • @nurfatiniafiqah1952
    @nurfatiniafiqah19528 жыл бұрын

    Good ... its helpful .. so we must memorize all the structures of amino acids? OMG ..

  • @anunucha
    @anunucha5 жыл бұрын

    what kind of protein doesn't start with methionine

  • @anyamiller514
    @anyamiller5146 жыл бұрын

    Why does Gly have a pk value? I wasn't aware that it had an ionizable group.

  • @anyamiller514

    @anyamiller514

    6 жыл бұрын

    Shouldn't it just be ignored in the calculation?

  • @TheBollywoodCritic
    @TheBollywoodCritic7 жыл бұрын

    May god bless you with a thousand sons.

  • @fukpoeslaw3613

    @fukpoeslaw3613

    6 жыл бұрын

    S.S .A Because we are not all feminists.

  • @leonardmulenga6021
    @leonardmulenga60212 жыл бұрын

    I still don't understand when it comes to the estimations How those where becoming neutral instead of +

  • @lixx1414
    @lixx14146 жыл бұрын

    QUESTION, SOMEONE HELP PLZ :D why are the pka=4.1 neutral with the guess of ph= 3.5 ?? shouldnt they both be positive...bc theyre above the guess??? leaving the over all charge to be +2?? help plz!

  • @lixx1414

    @lixx1414

    6 жыл бұрын

    +AKLECTURES

  • @lovelly2
    @lovelly24 жыл бұрын

    For problem 2, I don't understand why cysteine did not gain a positive charge. The pH is less than the pka which means it should have been protonated. I understand the charges for all the others but do not understand your explanation of cysteine.

  • @czechmeoutbabe1997
    @czechmeoutbabe19974 жыл бұрын

    I don't understand why sometimes he says the higher pKa means that it has a *positive charge* at a pH, but sometimes it has a *neutral* charge at that same pH value. They're both above the pH, I don't see the difference.

  • @pathbasics
    @pathbasics4 жыл бұрын

    So SH can't gain any more protons when its pka is above the ph? 12:35

  • @harinisuresh456
    @harinisuresh4564 жыл бұрын

    How come you have solution for all my assignments!!

  • @ACEF22able
    @ACEF22able8 жыл бұрын

    ....What about that pka = 8.3 at cys???

  • @soniproductionz

    @soniproductionz

    8 жыл бұрын

    +Anthony Chang At a pH of 7 it is protonated so it is S-H which has a neutral charge. Only after 8.3, it becomes deprotonated so it will be S- = -1 charge.

  • @user-mh1pu8gk1s
    @user-mh1pu8gk1s5 жыл бұрын

    想要中文字幕😭😭😭😭😭

  • @123Rukhsaar
    @123Rukhsaar8 жыл бұрын

    isn't it 2 similar points take an average of them in example 2 wouldn't it be (8.0+8.3)/2=8.15?

  • @soniproductionz

    @soniproductionz

    8 жыл бұрын

    +Rukhsaar A Close, it is similar points around the average pH at which the net charge is 0. In example 2 that was pH of 7 so similar points around that pH of 7 would be 8.0 and 4.1.

  • @ZippyandOllie
    @ZippyandOllie7 жыл бұрын

    wait, so why are the 4.1 neutral again?

  • @shanikhan00

    @shanikhan00

    7 жыл бұрын

    Because they were COO(-) to begin with. 4.1 makes it larger than the pH of 3.5, therefore they will gain an H and become COOH (neutral)