Embed Size (px)
Citation preview
chat with Fred and ChrisNov. 19th, 2010
• rhombohedra: SEM observation vs time, 24h-72h
• Bodega Marine Station payments - order Mytilus californianus
• spicule matrix proteins: produce in yeast (Liam Holt), then harvest and isolate with TAP (twice ).
• which proteins do we try?• can we go for the 5 most abundant in spicules?
• do we agree about the “protein-function” assay on calcite polished crystals? It only shows ACC-stabilization function.
• other, better ideas?
Mechanism of transformation of amorphous calcium carbonate into calcite in the sea urchin larval spiculeY Politi, RA Metzler, M Abrecht, B Gilbert, F Wilt, I Sagi, L Addadi, S Weiner, and PUPA GilbertProcs. Natl. Acad. Sci. USA 105, 17362-17366 (2008)
sea urchin spicules revealed 3 phases : 2 Amorphous Calcium Carbonate (ACC), and 1 biogenic calcite: type 1, type 2, type 3.
type 1type 2type 3?
spicule
shadow
Si wafer
X-PEEM (photoelectron emission microscopy) image and spectra
Transformation and crystallization energetics of synthetic and biogenic ACCAV Radha, TZ Forbes, CE Killian, PUPA Gilbert, and A NavrotskyProcs. Natl. Acad. Sci. USA, 107, 16438-16443, 2010.
transitions type 12 and type 23 are both energetically downhill
isothermal acid-solution calorimetry data
red = hydrated ACCgreen = anhydrous ACCblue = crystalline CaCO3
transitions type 12 and type 23 are both energetically downhill
isothermal acid-solution calorimetry data
red = hydrated ACCgreen = anhydrous ACCblue = crystalline CaCO3
Transformation and crystallization energetics of synthetic and biogenic ACCAV Radha, TZ Forbes, CE Killian, PUPA Gilbert, and A NavrotskyProcs. Natl. Acad. Sci. USA, 107, 16438-16443, 2010.
transitions type 12 and type 23 are both energetically downhill
isothermal acid-solution calorimetry data
red = hydrated ACCgreen = anhydrous ACCblue = crystalline CaCO3
Transformation and crystallization energetics of synthetic and biogenic ACCAV Radha, TZ Forbes, CE Killian, PUPA Gilbert, and A NavrotskyProcs. Natl. Acad. Sci. USA, 107, 16438-16443, 2010.
red = type 1, hydrated ACCgreen = type 2, anhydrous ACCblue = type 3, biogenic calcite
R+G=YellowR+B=MagentaG+B=Cyan
Component Analysis of X-PEEM data (X-ray PhotoElectron Emission spectroMicroscopy)
CaCO3 phase transitions in the sea urchin larval spiculeYUT Gong, CE Killian, IC Olson, NP Appathurai, RA Metzler, AL Amasino, FH Wilt, and PUPA Gilbertin preparation
type 1type 2type 3 sequence experimentally confirmed in spicule cross-section (20-nm pixels)
500 nm
type 12 and 23 are fundamentally different transitions
red = type 1, hydrated ACCgreen = type 2, anhydrous ACCblue = type 3, biogenic calcite
R+G+B=WhiteR+G=YellowG+B=CyanR+B=Magenta
spicule cross-section (20-nm pixels)Component Analysis of X-PEEM data(X-ray PhotoElectron Emission spectroMicroscopy)
CaCO3 phase transitions in the sea urchin larval spiculeYUT Gong, CE Killian, IC Olson, NP Appathurai, RA Metzler, AL Amasino, FH Wilt, and PUPA Gilbertin preparation
RGB enabled
red = type 1, hydrated ACCgreen = type 2, anhydrous ACCblue = type 3, biogenic calcite
simulation modelsRG inhibited
type 12 and 23 are fundamentally different transitions
CaCO3 phase transitions in the sea urchin larval spiculeYUT Gong, CE Killian, IC Olson, NP Appathurai, RA Metzler, AL Amasino, FH Wilt, and PUPA Gilbertin preparation
RGB enabled
1 µm
red = type 1, hydrated ACCgreen = type 2, anhydrous ACCblue = type 3, biogenic calcite
experimental data simulation models
RG inhibited
type 12 and 23 are fundamentally different transitions
type 12 transition is biologically controlledtype 23 transition is spontaneous
CaCO3 phase transitions in the sea urchin larval spiculeYUT Gong, CE Killian, IC Olson, NP Appathurai, RA Metzler, AL Amasino, FH Wilt, and PUPA Gilbertin preparation
RGB map
Green edge
Red edge
Magenta nanoparticles
Hydrated ACCAnhydrous ACCBiogenic calcite
red or green edges
magenta nanoparticles
Yutan: I see just as many cyan nanoparticles as magenta nanoparticles.What did you change here?are the above slides no longer valid?
48h and 72h spicules were extracted and deminineralized with acetic acid,neutralized with Tris buffer at pH 8, then dialyzed against DDH2O
sanity check:a droplet of DDH2O was deposited on a calcite crystal, then air-dried
sanity check:a droplet of DDH2O was deposited on a calcite crystal, then air-dried
re-crystallized calcite is more disordered:lower peaks 2 and 4
