Research output: Contribution to journal › Article › peer-review
Protein/Ice Interaction : High-Resolution Synchrotron X-ray Diffraction Differentiates Pharmaceutical Proteins from Lysozyme. / Bhatnagar, Bakul; Zakharov, Boris; Fisyuk, Alexander et al.
In: Journal of Physical Chemistry B, Vol. 123, No. 27, 17.06.2019, p. 5690-5699.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Protein/Ice Interaction
T2 - High-Resolution Synchrotron X-ray Diffraction Differentiates Pharmaceutical Proteins from Lysozyme
AU - Bhatnagar, Bakul
AU - Zakharov, Boris
AU - Fisyuk, Alexander
AU - Wen, Xin
AU - Karim, Fawziya
AU - Lee, Kimberly
AU - Seryotkin, Yurii
AU - Mogodi, Mashikoane
AU - Fitch, Andy
AU - Boldyreva, Elena
AU - Kostyuchenko, Anastasia
AU - Shalaev, Evgenyi
PY - 2019/6/17
Y1 - 2019/6/17
N2 - Protein/ice interactions are investigated by a novel method based on measuring the characteristic features of X-ray diffraction (XRD) patterns of hexagonal ice (Ih). Aqueous solutions of four proteins and other solutes are studied using high-resolution synchrotron XRD. Two pharmaceutical proteins, recombinant human albumin and monoclonal antibody (both at 100 mg/mL), have a pronounced effect on the properties of ice crystals, reducing the size of the Ih crystalline domains and increasing the microstrain. Lysozyme (100 mg/mL) and an antifreeze protein (1 mg/mL) have much weaker impact on Ih. Neither of the proteins studied exhibit preferred interactions with specific crystalline faces of Ih. It is proposed that the pharmaceutical proteins interact with ice crystals indirectly by accumulating in the quasi-liquid layer next to ice crystallization front, rather than directly, via a sorption on ice crystals. This is the first report, to the best of our knowledge, of major difference in the protein/ice interaction between non-antifreeze proteins. Another important finding is a detection of a second (minor) population of ice crystals, which is tentatively identified as a high-pressure form of ice, possibly IceIII or IceIX. This finding highlights a potential role of mechanical stresses in freeze-induced destabilization of proteins.
AB - Protein/ice interactions are investigated by a novel method based on measuring the characteristic features of X-ray diffraction (XRD) patterns of hexagonal ice (Ih). Aqueous solutions of four proteins and other solutes are studied using high-resolution synchrotron XRD. Two pharmaceutical proteins, recombinant human albumin and monoclonal antibody (both at 100 mg/mL), have a pronounced effect on the properties of ice crystals, reducing the size of the Ih crystalline domains and increasing the microstrain. Lysozyme (100 mg/mL) and an antifreeze protein (1 mg/mL) have much weaker impact on Ih. Neither of the proteins studied exhibit preferred interactions with specific crystalline faces of Ih. It is proposed that the pharmaceutical proteins interact with ice crystals indirectly by accumulating in the quasi-liquid layer next to ice crystallization front, rather than directly, via a sorption on ice crystals. This is the first report, to the best of our knowledge, of major difference in the protein/ice interaction between non-antifreeze proteins. Another important finding is a detection of a second (minor) population of ice crystals, which is tentatively identified as a high-pressure form of ice, possibly IceIII or IceIX. This finding highlights a potential role of mechanical stresses in freeze-induced destabilization of proteins.
KW - ICE RECRYSTALLIZATION
KW - ANTIFREEZE PROTEINS
KW - SOLUTE CRYSTALLIZATION
KW - STABILITY
KW - WATER
KW - ADSORPTION
KW - KINETICS
KW - BINDING
KW - PHASE
KW - LINE
UR - http://www.scopus.com/inward/record.url?scp=85069627446&partnerID=8YFLogxK
U2 - 10.1021/acs.jpcb.9b02443
DO - 10.1021/acs.jpcb.9b02443
M3 - Article
C2 - 31260313
AN - SCOPUS:85069627446
VL - 123
SP - 5690
EP - 5699
JO - Journal of Physical Chemistry B
JF - Journal of Physical Chemistry B
SN - 1520-6106
IS - 27
ER -
ID: 21047932