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Welcome to Cryo Talk,

a bite-sized bio podcast sponsored by Thermo Fisher Scientific,

featuring conversations between your host, Ava sson,

and experts in the field of cryo-electron microscopy.

Today on cryo talk

Today on cryo talk,

we are joined by Reinter Lab head in the Department of Microbiology at Monas

University.

He talks about how he determined the structure of an enzyme that turns air into

electricity. Yes,

It's about half a half a million hydrogen, um, in the air.

And bacteria use it as a auxiliary energy source when they starve,

when they have nothing else to live on.

And our cryo em helped reveal the details of this protein,

But it also told us the mechanism that the enzymes using to take the electrons,

take the energy from hydrogen,

and then direct them to put bacterial membrane into a thing called the electron

transport chain that allows the bacteria to make a t b.

He also shared some of his favorite places he discovered while traveling,

including Shark Bay,

The whole thing's a World Heritage area, and it's, it's this place where the,

the Red Desert of Inland Australia meets like tropical sea, right?

So you've got big sea grass meadows and Dugongs, and, um, coral reefs and,

and it's an incredible special place after

All in this episode of Cryo Talk.

Hi Anne, welcome to Cryo Talk. I'm Ava Amon,

and I'm here today with Reese Reese's lab head in the Department of Microbiology

at Monist University in Australia. So, hi Reese, how are you today?

Yeah, good. Yourself?

I'm good. Thanks for joining us. Um,

so we usually start off with, um, a little bit of background.

So why don't you share a bit of your career path?

How did you get to where you are?

Yeah, sure. I guess it's maybe a bit more convoluted than some people.

I did high school on an island called Kangaroo Island in South Australia,

in Australia, and then went to Adelaide, which is the state,

the capital of South Australia for undergraduate. Um,

but then I sort of took a break between honors and PhD.

We do honors in Australia first, and then we do PhD and,

and I lived to Japan for a year,

and then I traveled for a couple of years and ended up in Scotland,

Glasgow of all places where I started a PhD in, uh, microbiology.

And then the lab next door to me was doing structural biology,

so I got into X-ray crystalography, followed that through. Um,

after I finished my PhD 2015, I returned to Melbourne and I,

I've been at Monash University ever since.

Oh, well, we're definitely gonna be talking a bit more about your, your break,

your little research break, um, later on in the, in the episode.

Um, but when I was doing my background research on you,

I came across this intriguing headline from earlier this year,

scientists Discover Enzyme that turns air into electricity.

Can you tell us a bit about that?

Absolutely. Yeah. So this is a bit of a wild ride earlier in the year.

So this is a project I've probably been working on myself and Professor Chris

Greening, who's also at Monash University, um,

working on for maybe four or five years. And, and,

and Chris kind of discovered over the past decade that bacteria that live in,

in soils, they utilize the, the small amount of hydrogen in the air.

So there's about half a part million hydrogen, um, in the air,

and bacteria use it as a auxiliary energy source when they're starved,

when they have nothing else to live on.

And we really wanted to isolate the enzyme and,

and sort of figure out how it works. And so we worked,

it took us about four or five years where we managed to isolate,

purify and use cryo electron microscopy to determine the structure and basically

show using enzyme assays and structural biology. But yeah, this enzyme is,

is capable of, of taking that amount of hydrogen, half a part per billion,

A million, sorry. And converting it into electricity.

Wow. And what, what could that knowledge be used for?

Well, we're hoping to further develop the enzyme to start thinking about making,

um, fuel cells and devices that can use electricity either in,

in trace amounts because of the trace levels of hydrogen that are present in the

air or in larger amounts to, to power kind of, you know,

green electrical devices.

Well, that would be cool, and it would be very cool.

So you used cranium for this, as you mentioned. Um, why,

what was the advantage of cranium?

Yeah, it's a really good question. So, I mean,

I guess I started out doing crystallography and that's, that's what I learned.

And, and for a long time that was what I was doing, but this,

this, um, enzyme that we're working on was, it's very large, um,

800 kilodons in total, and it's, it attaches to the membrane,

um, and it also has a lot of flexibility and we also can't make very much of it.

And so it's just a,

it's the structural target that would be completely inaccessible to,

to crystallography. Um,

but the advantages that modern cryo em allowed us to basically freeze

grids and, and the first, um,

grids that we collected gave us nice high resolution data. So yeah,

it was a perfect technique for, for sort of studying this sample. So

What do you think, um,

the cryo OEM structure revealed that you would not have been able to see

otherwise?

Yeah, so, so I guess, um, we'd revealed the structure, the, the,

the structure of this enzyme complex that we wouldn't have been able to to see

by any other method. And, and this is an 800 kilo complex, and,

and it's also attached to the membrane and it's quite flexible.

So it was not a target that would be accessible by, by cryo yam, sorry,

by crystallography. And in fact,

we couldn't make enough of the protein to do crystallography.

So it revealed the structure, but also, um, we had, we had no idea what this,

this structure looked like. So it consists of eight copies of one subunit,

eight copies of another subunit, and four copies of the final subunit.

We didn't know how they went together. And so it told us that,

but it also told us the mechanism that the enzymes using to take the electrons,

take the energy from hydrogen and then direct them into the bacterial membrane

into a thing called the electron transport chain that allows the,

the bacteria to make attp.

Oh, that sounds like a, it sounds like it was really complicated,

like not even really knowing what you were looking for from the start.

Yeah, no, exactly. And so we used, we used a method I'm really a,

a fan of for protein purification these days,

which was just to chromosomally tag.

So we put an affinity tag on one of the genes of the complex in the chromosome

and then just pull it out from its host organism.

So microbacteria smoke is the host organism, and so we ended up isolating it.

Another subunit we didn't know was part of the complex and got all this rich

biological information that we wouldn't have got if we'd just made it more

traditionally in recombinant expression in, in a coline.

Yeah, that's cool. It's kind of like the,

the more realistic situation that if you just looked at one subunit and

Exactly. Exactly. And, and I think this is a, you know, this is a real power of,

of cryo em,

and you see more and more people successfully applying cryo tomography,

so looking at molecules in whole cells.

And so then you're getting the full context context,

and I think this is really where structured biology's gotta go and where it's

going, and it gives us a much more realistic picture of what's going on.

Yeah, it kind of brings me to my next question, which is, um,

what was it that initially drew you to cryo e m? Like,

do you remember when you first learned about the technique?

Yeah, I do actually.

So this was 2013 and I was getting pretty close to finishing my, um,

PhD in, in the uk. And, and there's, I think it was a, um,

CCP four is a initiative that has made a lot of crystalography software and they

have a,

a meeting every year where people go to and they discuss the latest methods.

And there was a talk where I think someone had done like a 3.5 and strong

resolution structure of like a ribosome by cryo em.

And that was the first time I'd seen anything remotely that resolution

that had been done by the technique. And so when I saw that, I was like, wow,

I mean, this is, it kind of blew me away, but at the same time I'm like, oh,

you know, everyone was saying, oh, it's only gonna be for big stuff.

It's only gonna be for, you know, a certain kind of targets. Mm-hmm.

And so I was watching it since then, and then over the next decade, I guess,

because that's all we're now, it's just got more and more and more advanced.

And for quite a while, Chris was still a better tool for my research, but, um,

Monash University established its electronic facility, I think about, um,

shortly after I ran about six or so years ago.

And a lot of my colleagues were doing really cool experiments,

producing really good data.

And so we just sort of started doing both in parallel and over time,

cryo em has just sort of, it's been an easier route and a and a,

a route that's allowed us to access a lot more complicated samples.

Yeah. And, and do you think, um,

over the past decade it's become more accessible? I mean,

I guess with people building new facilities, it must be right?

Yeah, absolutely. Initially in Australia,

Monash had the first one that was the only one.

So initially getting access to the instruments was,

was quite difficult because there's a lot of demand. But, um, there's,

there's at least two, um, facilities in, in Melbourne now,

and many other facilities around the country, which I think alleviate that.

The collection, um, has got a lot faster.

People have got a lot more skill dealing with manipulating samples.

And also the software is you, you know, very easy to use nowadays. So the,

the barrier for, for accessing and processing your data once you get it,

is lower now as well. So I think it's really maturing as a technique.

Yeah. Yeah, that's, I mean, we,

we're now at the end of season two of this podcast, and so from all the,

the stories that I've heard over the past couple of months,

it really sounds like, you know,

things are really picking up and people are using it so much more often and for

so many different things, it's, it's really interesting to hear. Um, yeah,

We, we did a, oh, sorry. We, we did a, um, a road test recently, well,

we had 75 kilo membrane protein that I wanted to crystallize with a ligands or

get the structure of a ligand.

And I'd already done a non ligand structure by crystallography a few years ago.

And so we, we did them side by side in the lab,

and we didn't manage to get a structure by crystallography,

but the first greens we sold,

we got a 2.3 and some structure by crow structured by crow with the ligand

bound. Um, wow. And so it's just, it was, it, it, it was the best technique.

We were trying them side by side and it was the best technique for the job.

So that was, it was really impressive.

Yeah, yeah. Yeah. So, so we just talked a bit about, you know,

your work from a few months ago and years ago,

but what are you working on at the moment? What's new?

Yeah, so I guess we're working on a few things. I feel like I've kind of,

I've sort of stamped, collected and,

and then not wanted to drop exciting projects. So the, the lab's major theme is,

so we're, we're microbiologists with bacteriologist and,

and we use structural biology and,

and biochemistry to sort of look at interesting systems and answer interesting

questions. So, for example, the hydrogenate,

these enzymes that are able to oxidize hydrogen from the air is,

is one branch of what we do. And another, um, a major interest is looking at,

uh, membrane transporters and bacteria,

especially pathogenic bacteria that they use to get iron and heme.

So iron sources from the host during infection.

So we're studying the structural biology and functional biology of those,

how they interact with, uh,

proteins like hemoglobin and how we could potentially inhibit them to prevent

bacteria from getting essential nutrients. So, so that's one other theme.

And the, the major other thing we're working on is, um,

looking at novel protein antibiotics, um,

to service alternatives to our dwindling supply of traditional antibiotics.

And so we've got a project where we're looking at a class of protein,

but antibiotics that inhibit, um, there's a really nice, uh,

multi protein complex in the outer membrane of gram-negative bacteria,

an

both to develop these compounds, these products specifically,

but also as a proof of principle for inhibiting this complexes antibiotic

target. And, and this project is, is getting off the ground,

but it's gonna feature crow quite heavily to do this massive MultiPro

membrane complex with the inhibitor bound.

And so we're really excited about that.

Yeah, yeah. I was just about to ask, are you still using cry omm for that?

But yeah, of course you are. Um,

so you mentioned it at the beginning, but you took a little bit of a break, um,

between undergraduate and PhD and it sounded really interesting.

And do you wanna tell us a bit more about what you did in that time and why you

came back to science? Yeah, sure.

I think, I think I always knew I wanted to come back to science.

I'd finished my honors and, and I'd been on Kangaroo Island,

which is a small island. There's about 5,000 people population.

And I went to Adelaide, which has about a million people.

So it was a much bigger city,

but I'd finished my honors and I probably could've stayed in Adela to do a PhD,

but I,

I felt there was a big wide world out there and I didn't wanna just get stuck in

this track. So I elected to kind of pause science for a while.

And the Japanese government, I think they still have it,

but there was a program called the Jet Program that basically takes you on a

government program, places you inside a, a school, a high school,

or a junior high school in Japan. So I applied for that program,

got on that program and, and got sent to Sapporo, um, which is the capital of,

of Edo in northern Japan. Mm-hmm. And I mean,

it was just like a new world for me.

Like I think I'd been overseas once at that stage and different culture,

different people, different place language,

and just really had a blast of a year. Um, the snow up there is amazing.

So snowboarding every weekend and, and not focusing on job,

but just focusing on kind of the experience and the life.

Yeah. And, um, did you know Japanese before you went?

No, I didn't know a word of Japanese before I went.

And it was easier 'cause I was in a city. Um, but I still, I, I took,

I think I took four or five hours of lessons a week, and so I was like solid,

intermediate by the time I left, but sadly,

it's all kind of slipped away subsequently.

Yeah. And, and I guess, did you,

did you learn anything from your break that has been helpful in your science

career after that?

I think so, yeah. So just to summarize briefly, so up to Japan,

I went to Northwestern Australia and lived in a place called Shark Bay for two

years. Um, animal themed names are apparently a thing that I like to do. Uh,

it was like a salt mine, so it was a mine that makes salt from the sea.

So I worked there as a chemist a couple of years, um,

and then traveled from Australia, well, I actually,

I took a plane from Australia to Bali,

but then I traveled from Bali to Scotland without flying, so

Oh wow. Went all through,

Uh, Southeast Asia and then ended up going across into China and across through

Kazakhstan, across the Caspian Sea and then through into Europe. And I mean,

I think that's really taught me about, I dunno,

different people and different cultures and,

and how kind of the world meshes together. And I mean,

how science is a fantastic career because,

because it's completely transferrable to many, many countries, right?

Mm-hmm. So I, I could said I could see myself living in multiple places,

but this was would be an option. I would've if I pursued a career in science.

Yeah. But yeah.

And then you decided you wanted to do your PhD. Um, what,

what was kind of, how did you know that your break was over?

I think there was a few things. So there was a point was,

was a process chemist in Western Australia. Mm-hmm.

And there was a point where like,

I should have been doing the process chemistry,

but there was like this really interesting side project where we were looking at

the composition of highly concentrated brains and I really wanted to do the

experimental side of the, more the operational side of it.

And so I think that that was the point where I was like, well,

I have to go back and do a PhD at a certain point. Yeah.

And then when I arrived in Glasgow, it was 2009, 2010,

so it was the middle of the global financial crisis. Mm-hmm.

And so there wasn't really any other jobs anyway to, to be, you know,

honest about it.

And so I thought it was a great time to do the PhD I'd always wanted to do, and,

and so I applied for some programs at University of Glasgow and was lucky enough

to get a, get a scholarship and things went from there.

Mm-hmm. Yeah. No, I, I finished my PhD 2008, so Yeah,

I know what you mean about there not being any jobs around that time. It's like,

yep. You just take what you can get. That's pretty rough.

You do what you can do. Um, yeah.

So a a question that I actually ask a lot of people is if,

do you have any favorite places that you've traveled to? So you've,

you've been in a lot of countries,

especially with the Overland journey from Bali to Scotland.

So any favorite places that you've come across along the way?

I think so many, to be honest with you, it's, it's really quite hard to pick.

So, um, kangaroo Island where I grew up is, it's,

it's fairly dry on the sometimes got quite a Mediterranean climate,

but the coasts beautiful sandy beaches, surfing great fishing,

really good diving actually.

So this is still a place that I consider really special. Um,

shark Bay where I was living, it's a whole thing's a world heritage area,

and it's, it's this place where the,

the red desert of Inland Australia meets like tropical sea, right? Mm-hmm.

So you've got big sea grass meadows and ju gongs and um, coral reefs and,

and it's an incredible special place. I was just back there, um,

a few weeks ago actually went up for, for a fishing trip, snowboarding Japan,

um, the Highlands of Scotland, like Tuscany and Italy, Istanbul, I don't know.

I mean, it's, it's impossible to, to to think to something and

They're all so different.

I just think the world's kind of a, you know, fantastically interesting place,

right? Like,

Yeah. Um, I've got a few quick fire questions for you.

Do you have a favorite book that you could recommend people?

It's a good question. Um, I think if, if I had to say,

you know, one piece of literature that I, I I would recommend,

I would say a hundred Years of Solitude by Gabrielle Gussy and Marque.

I think it's just really a beautiful piece of literature that to captures so

much of humanity and I'm, I'm reading less literature these days,

I should get back into it. So that would be,

that would be a book I would definitely recommend.

And, and what about, um, films or TV shows?

Do you have any favorite things that you can recommend?

Yeah, yeah. So I've, I've been kind of,

I've been watching less TV shows in cinema lately, um,

but still getting into a few things. But I was recently in the United States,

I went across to to California and I watched the entire series of Chernobyl from

start to finish on the plane and finish was just blown away. It, it was,

I mean, it was a captive audience and it's, it's a tough, I mean,

it's a tough watch, right? Like it's really, but yeah,

I had the time to appreciate it and it was, it was just this really slow burn.

I gotta the end of it, but my wife's from, from, from Eastern Europe,

from Serbia,

and I thought they captured something about the culture and the people so well

and, but just laid out this whole story. And so I would really recommend, um,

that as a series. Yeah.

Yeah. I still need to finish it. Um,

I started it during the pandemic and it was just a bit too much.

It was just a whole global pandemic going on and I'm watching this thing about

Chernobyl and I was like, I just can't deal with this right now, but yeah.

Should pick it up again. Yeah, I

Started, I started, I think I started man in the High Castle in the Pandemic,

and then I was like, thank you. So I totally get it.

Yeah. And do you like listening to

Music? Yeah, yeah. I was do a fair bit of music and,

and these days it's pretty eclectic in my, in my younger years,

I guess I was kind of a bit of a music sno.

I listen to a lot of like indie rock and I was like, oh, you can,

you can't listen to this, but you can listen to that. Um,

but in more recent times I've become completely agnostic and like,

anything that, that sounds good. Um, so yeah, a lot of eighties,

a lot of nineties, a lot of, um, yeah,

still quite alternative kind of indie rock like stuff like neutral milk hotel

and the Decemberists and um, the shins and, and things like that.

But also my daughter's five and she really likes, you know,

Katie Perry and she's getting Tell and John,

so I kind of follow along and I, I kind of, I don't regret,

but I think it was a,

it was a poor decision on my earlier selves part to be exclusionist about what

music was good to listen to and mm-hmm. If it makes you happy, then,

then you should listen to it.

Yeah. Yeah. It is fun to explore lots of different types of music too. This,

um, I mean,

we already talked a little bit about science versus other things,

but another of my quickfire questions is, if you were not a scientist,

what would you be?

No. Do I have to get paid to do it?

It's just an imaginary career. So

A man of pleasure, a traveling adventure, I dunno. No. Um,

so a few few things I could, I could see myself doing. I, I, I think I could,

I could probably cook for a living. I really enjoy cooking, so,

and there's a certain kind of parallel, I think,

to being in the kitchen and having that organization and, um,

and being a scientist or I think we're discussing it the other day,

and I think being a chef would be so much higher pressure.

It's like if an experiment fails in the lab, it doesn't matter.

Just do it again. If cooking, if it isn't right and the meal goes out,

that's bad, then, you know, there's, there's real world consequences. So yeah,

that would be high pressure.

Yeah, it's real people eating your food.

But aside from that, I mean, I, I think I might've enjoyed being an engineer.

Like it's nice being a scientist, but one thing that, you know,

you're, you're discovering new knowledge,

but whether or not that new knowledge is translated is sometimes not up to you.

Mm. And so maybe being at the other end of it where you're like,

you're taking established knowledge,

but you're trying to make something or maintain something that keeps society

running, I think I could be quite satisfied doing that.

Mm-hmm. Yeah. And you, and you mentioned you like cooking.

Do you have any favorite recipes, any favorite things you like to make?

My wife kind of ribs me because I don't like to repeat cooking things too,

too much, right? Mm-hmm. So it's like, it's,

everything's on a three monthly rotation, but so tonight I, um, I put a,

a lamb shoulder on this morning and so cooked it all day at a 20 degrees.

And so basically you put it on a bed of onions and then it just kind of cooks

down to tenderness. And then you take that and put it in, um,

tortillas with some Greek salad and, and some ssss. And this is delicious.

I think this is it for the, for the amount of effort it takes to cook.

I think it's, it's a winner for me.

Yeah, it takes a long time. You need to plan. You can't just decide last minute.

And, um, and then the,

the last question is always, uh,

an a difficult one for maybe or interesting one.

Is there any piece of advice that advice you've received during your career or

anything that you would want to pass on to scientists who are just starting out?

Yeah. Yeah. I mean, I think you have to be,

you have to be really interested and passionate about what you're doing. And,

and I think this was something gone, other podcasts said, but you,

you wanna wake up in the morning thinking about something and wanting do

something not because have

inevitably after

it's gonna really make you feel like this path is quite difficult.

But if you have the intelligence and the passion and the perseverance,

then there's a good chance you'll still get to where you get to.

So try not to let any individual set back, um,

just get you, get you down too much.

Mm-hmm. Mm. Good advice. Um,

so thank you Reese. Thanks so much.

It was really good catching up with you in today's podcast.

And that brings us to the end of our episode today.

So thank you everyone for listening to or watching cryo talk.