National Center for Earth & Space Science EducationStudent Spaceflight Experiments Program

Mission 3 to the International Space StationTitle A Study of How Microgravity Affects the Enzymes in Amyotrophic Lateral Sclerosis (ALS) Using the Model of Papain and Gelatin

Teachers in Space, Inc./Space Frontier Foundation CommunityCo-Principal Investigators: Robert Edmiston, Jaclyn Martin, Luke Redito, Harshit Saini, and Sanju VardhanCo-Investigators: Genna Owen, Carissa Sage, and Hannah SchroeterCollaborators: Rashad Abdulla, Varun Bansal, Caleb Bryant, Olivia Escandell, Dave Foster, Anuj Mehindru, and Krunal Patel

Inspiration for the Experiment• The researchers wanted to

better understand ALS and raise awareness of the disease when their teacher, Jason Whitworth, was diagnosed in the fall of 2011.

• The team of student researchers refers to the experiment as Project Whitworth.

Project WhitworthBackground on glutamate

• Glutamate is an amino acid that acts as a neurotransmitter.

• The amount of glutamate that humans need is miniscule and is already manufactured by the body or obtained in whole foods.

• In the synaptic cleft between motor neurons, glutamate is normally deactivated and recycled by enzymes.

Project WhitworthBackground on ALS patients and glutamate

• In ALS patients, glutamate does not get broken down, but builds to toxic levels, and eventually causes the neurons to die.

• As the neurons die, ALS patients lose control of voluntary muscles.

Project WhitworthBackground information on enzymes and proteins

• Enzymes work by pairing with specific substrates at an active site, often referred to as a “lock and key.”

• When an enzyme and substrate bind together, the enzyme changes shape, often referred to as an “induced fit,” and allows the enzyme and substrate to function together.

• The enzyme can then break down the large protein polymer into smaller amino acid subunits.

Project WhitworthOur hypothesis

• The researchers’ hypothesis was that the enzyme would be able to function more effectively in a microgravity environment.

• We based this on the assumption that, with less gravity, there would be fewer restrictions during the enzyme-protein pairing and shape change.

Experiment ProceduresFluid Mixing Enclosure (FME) – Type 3

• To initiate the reaction, Clamp A was released and the FME was shaken vigorously for 20 seconds.

• To terminate the reaction, Clamp B was released and the FME was shaken vigorously for 20 seconds.

0.6 ml Powdered Papain

5.5 ml Gelatin

0.1 ml Ascorbic Acid

Project Whitworth LaunchLaunched January 9, 2014

from Mid-Atlantic Regional Spaceport, Wallops Island, VAon an Orbital Sciences Antares rocket & delivered by the Cygnus ferry vehicle

Project Whitworth at Launch Controlsent to Coach Whitworth from NASA

Thumbs up from the Launch Control Center Chris Scolese - Director, Goddard Space Flight Center

Dave Thompson - Orbital Chairman, President and CEO Frank Culbertson - Orbital, Executive Vice President and General Manager Dale Nash - Executive Director, Virginia Commercial Space Flight Authority

Chris DeMars - Orbital, Manager for FAA and FCC Licensures Michael Suffredini - ISS Program Manager

Bill Wrobel - Director, Wallops Flight Facility

Project Whitworth on the ISSExperiment initiated by Mike Hopkins March 5, 2014

Experiment terminated by Koichi Wakata March 8, 2014Total Run Time: 3 days, 3 hours, 59 minutes

Mike Hopkins and Koichi Wakata performing experiments on ISS

Project Whitworth on the ISSJanuary 12 – March 10, 2014

Experiment run March 5 – March 8

Two Facebook message shout-outs sent to Ms. McCormick from

Astronaut Rick Mastracchio (ISS Expedition 38):“Tell Jason we said hello from the International Space Station.”

(March 1, 2014 at 3:02 pm)

“It seemed to go well. Thanks for sending us science.”(March 7, 2014 at 5:29 pm)

Project Whitworth on EarthExperiment initiated on March 7, 2014

Experiment terminated on March 10, 2014Total Run Time: 3 days, 3 hours, 59 minutes

Back to EarthThe experiment was returned to Kazakhstan onboard Soyuz TMA-10M with Expedition 38 on March 11, 2014

Flew with Commander Oleg Kotov, Mike Hopkins, and Sergey Ryazanskiy

Back to Houston then to FloridaReturn to Melbourne on March 13, 2014 where we opened it

with Brevard ALS Support Group

Project WhitworthData Analysis

• The team performed a Biuret protein assay to determine the levels of protein remaining after the enzyme reaction.

Project WhitworthData Analysis

• Researchers used a spectrophotometer to determine the amount of remaining protein by measuring the absorption levels for each sample.

Project Whitworth DataFME Run by Amount

PapainAmountGelatin

Amount Ascorbic

Acid

Wavelength AbsorbanceTrial 1

AbsorbanceTrial 2

ISS Mike Hopkins

0.6 g 5.2 mL 0.1 g 540 nm 1.630 1.915

Official #1

GennaOwen

0.6 g 5.2 mL 0.1 g 540 nm 2.998 3.034

Official #2

CarissaSage

0.6 g 5.2 mL 0.1 g 540 nm 3.093 3.087

Backup #1

Hannah Schroeter

0.6 g 5.2 mL 0.1 g 540 nm 3.054 3.059

Backup #2

RickFleming

0.6 g 5.2 mL 0.1 g 540 nm 3.046 3.044

Spectrophotometer Data for FME Samples

FME Samples

Abs

orba

nce

Leve

ls

Trial #1

Trial #2

ISS Official #1 Official #2 Backup #1 Backup #2

1.630

1.915

2.998 3.0343.093 3.087 3.054 3.059 3.046 3.044

Project WhitworthConclusion

• According to ANOVA statistical analysis, the difference between the two groups was significant (the ISS sample was ~40% more efficient at breaking down the protein than the samples here on Earth).

• This supports our original hypothesis that the enzymatic breakdown of proteins would be more efficient in a microgravity environment compared to that on Earth.

Thank You! We would like to express our gratitude to the following:

AeroInstitute National Institutes of HealthAirship Ventures NanoRacks, LLCALS Association Florida Chapter National Aeronautics and Space AdministrationApogee Books NCESSEBoeing Company Palmdale HotelBrevard ALS Support Group Paralyzed Veterans of AmericaCalifornia Near Space Project Skylark Soaring Celestial Circuits Smithsonian National Air and Space MuseumColumbus Space Program Student Spaceflight Experiments Program Embry-Riddle Aeronautical University Space Frontier FoundationFederal Aviation Administration Space Portal at the NASA Research Park Florida Today SpaceXJohns Hopkins University Teachers in Space, Inc.Masten Aerospace XCOR AerospaceMojave Air & Space Port zero2infinity