Abstract
Laser-induced microfragmentation of LB nanotemplate-induced protein crystals in glycerol solution results in distinct, coherently diffracting domains. Only crystals produced according to the Langmuir-Blodgett (LB) nanotemplate technique reveal in all four proteins being tested (lysozyme, insulin, thaumatin and ribonuclease) domains highly radiation resistant, while the crystals produced by the standard hanging drop crystallization method do not. Actually the very same laser exposure causes the disappearance of these "classical" protein crystals during the same time frame of 40 min needed for the laser cutting in all four proteins being tested. The microdiffraction of microcrystals prepered by the combination of Langmuir-Blodgett and Laser technologies proves that not only the Lysozyme survives the process, as shown recently by nanodifraction, but also all three other model proteins appear to behave similarly well, namely insulin, thaumatin and ribonuclease. The result confirms the emerging of a new biophysical technique uniquely usefull for synchrotron radiation studies based on small protein microcrystals uniquely radiation resistant when prepered by LB nanotemplate and subsequently fragmented by Laser.
Highlights
Laser-microdissection have recently (Pechkova et al, 2013; Pechkova and Nicolini, 2010) been successfully used to dissect Langmuir-Blodgett lysozyme crystals
Crystals produced according to the Langmuir-Blodgett (LB) nanotemplate technique reveal in all four proteins being tested domains highly radiation resistant, while the crystals produced by the standard hanging drop crystallization method do not
The result confirms the emerging of a new biophysical technique uniquely usefull for synchrotron radiation studies based on small protein microcrystals uniquely radiation resistant when prepered by LB nanotemplate and subsequently fragmented by Laser
Summary
Laser-microdissection have recently (Pechkova et al, 2013; Pechkova and Nicolini, 2010) been successfully used to dissect Langmuir-Blodgett lysozyme crystals Claudio Nicolini et al / American Journal of Biochemistry and Biotechnology 10 (1): 22-30, 2014 general extension to protein crystallography (Nave, 1999; Pechkova and Nicolini, 2004). This is thereby the main objective of this comunication. We have used the term laser-microdissection for the cutting of a crystal by a laser beam into smaller pieces (Pechkova et al, 2013) while microfragmentation is used for the separation of a microdissected crystal into smaller fragments due to effects such as cavitations at domain boundaries and solvent interpenetration.We will explore in this contribution the question whether protein crystals differing in perfection and X-ray radiation stability differ in microdissection and microfragmentation behavior. Atomic Force Microscopy (AFM) suggests differences in surface topologies for protein crystals grown according to the two methods (Santucci et al, 2011)
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