Otwarty dostęp

Effect of macromolecular mass transport in microgravity protein crystallization

Arayik Martirosyan
Arayik Martirosyan
, 
Lawrence J. DeLucas
Lawrence J. DeLucas
, 
Christina Schmidt
Christina Schmidt
, 
Markus Perbandt
Markus Perbandt
, 
Deborah McCombs
Deborah McCombs
, 
Martin Cox
Martin Cox
, 
Christopher Radka
Christopher Radka
 oraz 
Christian Betzel
Christian Betzel
   | 10 wrz 2019
Gravitational and Space Research's Cover Image
O artykule
Poprzedni artykuł
Następny artykuł
Zacytuj
Article Category: Research Note
Data publikacji: 10 wrz 2019
Zakres stron: 33 - 44
DOI: https://doi.org/10.2478/gsr-2019-0005
Słowa kluczowe
© 2019 Arayik Martirosyan, Lawrence J. DeLucas, Christina Schmidt, Markus Perbandt, Deborah McCombs, Martin Cox, Christopher Radka, Christian Betzel published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

Hardware and capillary design for LMM Biophysics-1 protein crystallization experiments. (A) Representative images of flight cassette with eight capillaries. The left image shows a cassette with eight capillaries. The image on the right shows a cassette with aligned and fastened top cover. (B) An empty capillary. (C) Schematic diagram illustrating the capillary shape and coordinates.
Hardware and capillary design for LMM Biophysics-1 protein crystallization experiments. (A) Representative images of flight cassette with eight capillaries. The left image shows a cassette with eight capillaries. The image on the right shows a cassette with aligned and fastened top cover. (B) An empty capillary. (C) Schematic diagram illustrating the capillary shape and coordinates.

Figure 2

Protein crystallization in microgravity applying the counter diffusion technique in the capillary. Images were recorded using the ISS LMM microscope, 2.5´ objective. (A) Lysozyme crystals. Images were recorded at t = 8725 min after sample thawing. (B) PfGST crystals. Images were recorded at t = 8674 min after sample thawing.
Protein crystallization in microgravity applying the counter diffusion technique in the capillary. Images were recorded using the ISS LMM microscope, 2.5´ objective. (A) Lysozyme crystals. Images were recorded at t = 8725 min after sample thawing. (B) PfGST crystals. Images were recorded at t = 8674 min after sample thawing.

Figure 3

Growth of lysozyme crystals in microgravity. Time-lapse images were recorded for three different areas (A, B, C) in capillary using LMM microscope with 2.5´ magnification. The imaging time extended to t = 8725 min, first image was taken t = 253 min after sample thawing.
Growth of lysozyme crystals in microgravity. Time-lapse images were recorded for three different areas (A, B, C) in capillary using LMM microscope with 2.5´ magnification. The imaging time extended to t = 8725 min, first image was taken t = 253 min after sample thawing.

Figure 4

Growth of PfGST crystals in microgravity. Time-lapse images were recorded for three different areas (A, B, C) in capillary using LMM microscope with 2.5´ magnification. The imaging time extended to t = 8674 min, first image was taken t = 301 min after sample thawing.
Growth of PfGST crystals in microgravity. Time-lapse images were recorded for three different areas (A, B, C) in capillary using LMM microscope with 2.5´ magnification. The imaging time extended to t = 8674 min, first image was taken t = 301 min after sample thawing.

Figure 5

Comparative growth rate and size distribution values (the length of major axis) of crystals along the capillary. Different regions (areas) along the capillary were measured from the protein end of the capillary toward the protein–precipitant interface. Error bars represent standard deviations of the average. Significance within a 99% confidence interval was determined using the Student’s t-test and is denoted by asterisk (*=p < 0.01). (A) Average growth rates with standard deviations of lysozyme crystals with lengths of major axis < 350 mm and > 350 mm. The sample sizes for crystals < 350 mm and > 350 mm are n = 6 and n = 9, respectively. (B) Average growth rates with standard deviations of PfGST crystals with lengths of major axis < 200 mm and 300–400 mm. The sample sizes for crystals < 200 mm and 300–400 mm are n = 3 and n = 3, respectively. (C) Average size distribution of lysozyme crystals for the areas from the capillary end 13–25 mm and 25–44 mm. The sample sizes for crystals in the areas 13–25 mm and 25–44 mm are n = 24 and n = 15, respectively. (D) Average size distribution of PfGST crystals for the areas from the capillary end 26–29 mm, 29–35 mm, 35–41 mm, and 41–44 mm. The sample sizes for crystals in the areas 26–29 mm, 29–35 mm, 35–41 mm, and 41–44 mm are n = 15, n = 20, n = 18, and n = 8, respectively.
Comparative growth rate and size distribution values (the length of major axis) of crystals along the capillary. Different regions (areas) along the capillary were measured from the protein end of the capillary toward the protein–precipitant interface. Error bars represent standard deviations of the average. Significance within a 99% confidence interval was determined using the Student’s t-test and is denoted by asterisk (*=p < 0.01). (A) Average growth rates with standard deviations of lysozyme crystals with lengths of major axis < 350 mm and > 350 mm. The sample sizes for crystals < 350 mm and > 350 mm are n = 6 and n = 9, respectively. (B) Average growth rates with standard deviations of PfGST crystals with lengths of major axis < 200 mm and 300–400 mm. The sample sizes for crystals < 200 mm and 300–400 mm are n = 3 and n = 3, respectively. (C) Average size distribution of lysozyme crystals for the areas from the capillary end 13–25 mm and 25–44 mm. The sample sizes for crystals in the areas 13–25 mm and 25–44 mm are n = 24 and n = 15, respectively. (D) Average size distribution of PfGST crystals for the areas from the capillary end 26–29 mm, 29–35 mm, 35–41 mm, and 41–44 mm. The sample sizes for crystals in the areas 26–29 mm, 29–35 mm, 35–41 mm, and 41–44 mm are n = 15, n = 20, n = 18, and n = 8, respectively.

Figure 6

Crystals of PfGST and lysozyme with additionally fluorescence-labeled aggregates. (A) Crystals of PfGST dimer with additionally fluorescence-labeled (Alexa Fluor 488) PfGST aggregates. Crystal images of PfGST were recorded on ISS using the LMM microscope, 2.5´ objective, with the FITC fluorescence filter. Images were recorded at t = 8742 min after sample thawing. (B) Crystal images of PfGST were recorded using a confocal laser microscope, 10´ objective, with the FITC filter (C) Crystal images of lysozyme monomer with additionally fluorescence-labeled (Alexa Fluor 594) lysozyme aggregates were recorded using the confocal laser microscope, 10´ objective, with the Texas Red filter. Images (B) and (C) were recorded after return of the samples on the earth.
Crystals of PfGST and lysozyme with additionally fluorescence-labeled aggregates. (A) Crystals of PfGST dimer with additionally fluorescence-labeled (Alexa Fluor 488) PfGST aggregates. Crystal images of PfGST were recorded on ISS using the LMM microscope, 2.5´ objective, with the FITC fluorescence filter. Images were recorded at t = 8742 min after sample thawing. (B) Crystal images of PfGST were recorded using a confocal laser microscope, 10´ objective, with the FITC filter (C) Crystal images of lysozyme monomer with additionally fluorescence-labeled (Alexa Fluor 594) lysozyme aggregates were recorded using the confocal laser microscope, 10´ objective, with the Texas Red filter. Images (B) and (C) were recorded after return of the samples on the earth.
eISSN:
2332-7774
Język:
Angielski
Częstotliwość wydawania:
2 razy w roku
Dziedziny czasopisma:
Life Sciences, other, Materials Sciences, Physics
Kanał RSS czasopisma