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[Music]

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[Applause]

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hello everyone so yes my name is Maureen

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Franco Pinter and I'm a NASA

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postdoctoral fellow can you hear me okay

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working with dr. Williams dr. hood and

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dr. Grover at Georgia Tech I'm also a

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proud member of the Center for chemical

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evolution and the Center for origins of

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life at Georgia Tech and today I will

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tell you about chemical mutualism

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between RNA and cationic lepsy public

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but first let's start with a quick

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definition of what do you mean by

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mutualism I'm sure I'll follow if all of

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you are familiar with the definition of

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mutualism in biology in which we have

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interactions in which both species

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benefit in the level of the organisms

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for instance we know that the bee

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pollinates the flower and the flower

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supplies food for the bee and it's the

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same thing when we talk about chemical

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mutualism in which we have interactions

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between species in the form of molecules

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in which both types of polymers benefit

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here we see this well the most complex

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molecular machinery that we know today

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the ribosome which is composed of both

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RNA and proteins we know that RNA makes

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proteins and proteins make RNA and we

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think that these types of interactions

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between different types of polymers were

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really important very early on in

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chemical evolution before diving to our

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mutualism project when we look at the

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formation of peptides on a prebiotic

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earth we face several challenges with

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condensation of amino acids into

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polypeptides the first challenge is

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comes from thermodynamics it is unfair

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unfavorable thermodynamically

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to form the amide bond in aqueous

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solutions next we have to kinetic

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barriers first to form the amide bonds

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it requires high activation energy

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barriers and second if we did form a

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dipeptide

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it can undergo another imitation to form

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this cyclic six membered ring which is

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called aikido piperazine which is

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hydrolytically very stable and so it's

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hard to further elongate

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polymers that are being formed to

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overcome these challenges or to

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polymerize peptide several solutions

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have been proposed for instance for the

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thermodynamic barrier we can simply

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shift to dry down reactions in which the

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thermodynamic equilibrium is change and

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now we can we can favour condensation

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dehydration reactions for the two

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kinetic barriers shown here we can use

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high temperatures but that could also

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lead to some decomposition of the amino

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acids themselves or we can use high

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energy molecules such as activating

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amino acids but there is a questionable

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a questionable availability of those

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molecules on a pre-wrath in the Center

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for chemical evolution we've come up

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with another solution and briefly as I

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mentioned polymerization of amino acids

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to form a polypeptide requires a high

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energy high activation energy barrier so

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a dry down we would need high

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temperatures but doctor had suggested to

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use these similar building blocks to

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amino acids so these are hydroxy acids

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which have alcohol instead of diamine

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glycolic acid for instance is the

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hydroxy acid analogue of glycine and

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lactic acid is the hydroxy acid analogue

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of alanine and we know that the atraxi

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acids were very readily abundant

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probably on the prebiotic earth because

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we see them in meteorites they come from

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the same type of synthesis as the amino

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acids and so they are Co localized

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together and we know that to form these

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polyesters so this step is pretty easy

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because it requires lower activation

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energy barriers so we can easily form

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polyesters but what will happen if we

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will mix these two types of building

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blocks amino acids and Roxy acids

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together so what we see under a dry run

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reactions is that we can form the aBSI

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peptides which are core polymers of

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amino nitrox acids or esters and amides

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and the reason why we can form these are

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these peptide bonds is through a

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mechanism

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called estimate exchange once we have

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our esters we have an activated carbon

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eel that can undergo a nucleophilic and

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chlorophyll attack with an amine in a

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process called experiment exchange and

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now we formed an amide bond and replace

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an ester bond so this process is very we

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can readily form that's it baptized just

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via dry down reactions so for the

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mutualism project our hypothesis is in

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interactions between different types of

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polymers were really important to shape

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chemical evolution and we want to in

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this project we want to establish that

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specifically interactions between

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cationic dupsy pathways in RNA we're

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really important and allowed them to

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mutually interactive promote the

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synthesis stability and function as

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model interactions we chose

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electrostatic interactions so we study

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interactions between cationic Li of

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between cationic Pepsi peptides and the

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negatively charged RNA backbone and we

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asked two main questions first we asked

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can we form Karianna cropsy peptides on

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VA Dryden reactions when we make these

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prebiotic environments the second

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question was if we did form carry on

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except see peptides can they mutually

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interact with RNA and promote it and in

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a way that will be really interesting to

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look at in terms of function structure

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and so on for the first question just

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for this slide please no photos because

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the paper has just been accepted to PNAS

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it's under embargo so the first question

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can we form cationic deficit better than

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prebiotic earth we formed they are will

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form deficit baptized via dry down

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reactions of hydroxy acids so we have

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lactic acid and glycolic acid and we use

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six different cationic amino acids three

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of them are the proteinaceous amino

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acids by that I mean the amino acid the

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quraíanic amino acids that are found

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today in

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proteins and they are incorporated

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during translation these are arginine

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histidine analyzing the three on the

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bottom here are shorter versions of

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lysing which have fewer methylene chains

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or methylene groups on the side chain

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these are only themed to Fordham you

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know Peter casted into three diamond or

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propionic acid these non proteinaceous

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amino acids are considered to be more

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prevalent lyrically plausible we find

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them in meteorites and in model

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prebiotic reactions so what we did in

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our experiments we dried down one I'd

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Roxie acid with either one of separately

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with either one of these different amino

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acids and we formed FC peptides now you

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can imagine how complex these different

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combinations will be because Arkady's

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cationic amino acids can link through

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different groups for instance when we

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dried our glycolic acid with lysing so

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license has two amino groups the Alpha

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amine and the epsilon Amin

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so both of these amines can potentially

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react them to form an amide bond we've

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characterized the deficit baptized using

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a variety of methods including FTIR mass

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spec h2 HPLC and so on but the method

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that was most informative was NMR and

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what it did it allowed us to look at the

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collective properties of the in-sample

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of molecules or the chemical signature

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of the Pepsi peptide that we form and

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what we found is very interesting so for

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instance in the case of lysing lysine as

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I mentioned can emulate just through the

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Alpha mean or just through the epsilon

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amine or using both the Alpha and the

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epsilon I mean what we found by NMR is

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that actually most of the polymers that

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we formed were worried those in which

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lies in a mediator the Alpha amine but

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still had a free epsilon amine and we

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have a pretty good understanding to why

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this is but in the beginning it was

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quite very well still it's really

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interesting too

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so 77 percent of all of our polymers

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look like that which implies it's just

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like in today's biology where the lysing

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will we have these I guess linear

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polymers with free amine groups on the

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cationic amino acids and hopefully you

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can see this work soon but just to

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summarize we have seen something very

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interesting is that the proteinaceous

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amino acids condense more extensively

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and selectively through their alpha mean

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compared to the non proteinaceous amino

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acids and we think that we understand

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now the chemical basis for selection of

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today's cationic amino acids over non

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proteinaceous amino acids that we also

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had in our soup so yes we can form

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cationic Pepsi peptides but what can we

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do with them can they take part in a

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mutualistic manner with our name so to

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that and dr. Lehman synthesized a

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cationic Pepsi peptide or a peptide and

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we had in our system both labeled RNA

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this was a u xx and a family both

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peptide or dab sleep at that and the

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first thing that we wanted to do is can

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we see physical association between the

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two types of polymers so we use the

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commonly used been shift a saying very

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briefly under native gel conditions when

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we run our na it will migrate to a

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certain extent but upon interaction for

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instance with a peptide it will Margaret

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a bit slowly and we should be able to

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man to look at that so what you can see

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here is a indeed where physical

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association between peptides and the

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cationic baptized and FC peptides the

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unbound RNA is labeled in red and we can

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see this smear and even the distinct

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band in orange that implies the

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colocalization of both the RNA and the

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peptide or dab see better once we've had

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this physical association we went to

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explore different ways or means in which

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we can look at the mutualism between the

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RNA and cationic CC peptides both

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synthesized FC peptides but also the

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ones is reformed

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dried-on reactions what we see here is

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one example of such a mutualism aspect

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that we saw which is really the most

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fundamental element of mutualism if

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interactions between different types of

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polymers if we have these interactions

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this should lead to a slower degradation

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rate of the polymers compared to if they

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are not associated and this is what we

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see here so we have our cationic Pepsi

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peptide with just one ester we can

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follow the degradation of this Esther

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because Esther is degrade much faster

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compared to Edmund bonds so looking at

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this HPLC chromatogram this here you can

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see the degradation of this dubsy

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peptide from one minute to 180 minutes

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the new peak that arises here is the

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segregate degraded deputy peptides so

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you can see the degradation this is

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without RNA once we introduce RNA to the

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sorry to the system you can see Arbus

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difference in the degradation rate so we

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introduce a tenma RNA duplex and you can

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see that there is barely degradation of

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that starting the app c-peptide when we

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go and quantify these differences here

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you can see the percentage of the intact

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dep c-peptide

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versus time and you can see in black

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without RNA in blue with RNA we can see

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that RNA duplex increases the the ab c--

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peptide half-life by about 30 fold so

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this is busting through robust

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protection to conclude we have seen that

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carry on who have shown that carry on

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adapt CPAP des can be formed via dry

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down reactions very robustly selectively

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and it was great to see that in addition

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with I've shown you that irony protects

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the esters within cationic Dempsey

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peptides and we go about and look at the

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different mutualism to Quincy and one

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more example that we saw is that I

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didn't show you is that cationic lipids

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can increase the thermal stability of

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our name and overall these findings

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suggested indeed cationic the cpap dates

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in our name

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interact to promote each other a

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structure and function they protect each

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other and the takeaway message from this

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lecture is that we think that yes

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interactions between different types of

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polymers was very very important in

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shaping chemical evolution because

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together they are going to be more

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stable and thus they will be selected

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further on and lastly I would like to

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think the Piz I worked with dr. Williams

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dr. huh dr. Grover our research groups

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here listed all of the co-authors so

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you're a team of researchers who work on

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this this project that I showed you

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specifically the leaders of this project

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are dr. Williams and dr. Weiman J

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heinous did the gel shift assay that I

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showed you Martin C helped with the NMR

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analysis dr. Lehmann synthesized

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cationic Pepsi peptides and looked at

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the degradation I say that I showed you

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of the deficit baptized and I would also

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like to thank the funding agency NASA

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postdoctoral program for funding me and

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also NSF NASA for their various grants

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and centers that they allow us to be at

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all right we have time for one quick

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hmm yes so potentially there there could

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be such Association I mean such an

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effect we also looked at the Association

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not through these labeled peptides and

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FC peptide I showed but also using just

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the labeled RNA and we still saw these

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smears just we couldn't see the

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colocalization but yeah I mean there

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could be some effect but it's not just

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30 the floor for all right I think we're