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 -