NASA-PAIR/ Proteomics Projects

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NASA-PAIR/ Proteomics Projects

• Crystal Austin
• Gerardo Lopez
• Elham Sarabi

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Interesting Info about Sea Urchin

• About 10 cm across and the spines are about 2 cm
• The spines are used for protection, movement and
  for trapping drifting algae
• They move surprisingly fast on their tube feet
  and spines
• They can also re-grow broken spines
• Used in public aquariums as an indicator for
  water quality

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Habitat and Distribution

• Live only in the ocean can not survive in fresh
  water. They are found from the intertidal to the
  deep ocean.
• Besides the S. California Coast,

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Location on the Food Chain

• Primary Food- Diet consists of algae, plankton,
  periwinkles, and mussels
• Predators- they are eaten by crabs, sunflower
  stars, snails, sea otters, some birds, fish and
  even people.

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Reasons Why We Study the Vitelline Envelope

• Location on egg?
• What is the vitelline envelope?
• Significance in fertilization
• Significance in research

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Outline
I. Question Being Asked II. Data Collection III.
Choosing the Best Model IV. Predicting the
molecular weights of the unknown data sets V.
Results Obtained
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Questions Being Asked?
Does the Vitelline Envelopes polypeptides from
two species of sea urchins have the same
molecular weight? Using the mechanically
isolated Vitelline Envelope in one species of sea
urchin, are the two chemical methods giving the
same results with respect to the number and size
of polypeptides?
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Purpose
TO DETERMINE THE MOLECULAR WEIGHTS OF UNKNOWN
PROTEIN BANDS IN A GEL BY USING STATISTICAL
MODELS.
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Comparing Strongylocentrotus Purpuratus to
Lytechinus Pictus

• File Studied
• S.purp, L.pictus, DTT treatment, 15 gel
• How DTT treatment works.

OH OH
HS-CH2-CH-CH-CH2-SH

• Goal
• Analyze file for comparisons.

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Comparing Chemical Treatments w/ a Manual Method

• File Studied
• Sea urchin VE removal
• Methods of isolating the polypeptides
• DTT
• Alpha-amylase

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Comparing Chemical w/ a Manual Cont...

• Manual (standard)
• Goal
• Analyze file for comparison.
• Find the relationship between the two chemical
  treatments and the manual method.

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Methods Used

• Using the 15 gel sample and the 12 gel sample,
  we approximated the dye front for each based on
  the end of the gel readings.
• We then isolated each lane on a new sheet and
  read the cm migrated for each band. (In order to
  get the best result, we adjusted the brightness
  and contrast in adobe photoshop)
• For each lane we ran three trials, then averaged
  for best results.
• We recorded all data on excel for future
  calculations.

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Methods Continued

• Using the recorded dye front and measured values,
  we calculated the relative mobility by dividing
  the cm migrated by the centimeters to the dye
  front.
• Using the standard, we calculated the best fit
  model with linear, quadratic, cubic, 4th, and
  some non-standard functions

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Modeling

• Looking for the best fitting model y
  abx y abxcx2 y abxcx2dx3
  y abxcx2dx3ex4 Nonlinear y
  abxcLNx
• Choosing the best model

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In Concluson...
The 12 gel sample contain less errors in fitting
a model, than the 15 gel sample. After choosing
a nonlinear standard model, it was found that the
natural log yeilded the smallest standard
deviation. It also maximized the degrees of
freedom overall, allowing us to have a more
normal distribution.
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References

• http//www.yorvic.york.ac.uk/projects/2/2.2.3.1.ht
  m
• http//www.sidwell.edu/sidwell.resources/bio/Virtu
  alLB/sea.html
• http//stanford.edu/group/Urchin/nathistory.html
• http//www.wcaslab.com/tech/Dithiothreitol.htm
• Proteomics Dr. Edward J. Carroll, JR.
• Data Analysis Dr. Larry Clevenson

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THANK YOU!!!!!!!
We Thank Dr. Carroll, Dr. Clevenson, Dr. Shubin,
Vred, and our fellow students for a great
CSUN/JPL- PAIR Program!
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THE END!