Langmuir–Blodgett nanotemplates for protein crystallography

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Alternative approaches to solve the complexity of protein function in the proteomic field required new arrays in nano-micro-scale dimension. Anodic Porous Alumina (APA), a geometrically ordered inorganic oxide, grown from ultrapure aluminum by anodic oxidation, was used in this work as an ordered array for the confinement of protein molecules by spin-coating technique. This microor nanostructured material can be used in different ways, both for the traditional fluorescence approach and for novel label-free methods. The aim of the present study is the proof of principle that the protein molecules can enter the APA micropores, as shown by fluorescein-labeled lysozyme confined into the APA template. Fluorescence intensity measurement as well as Raman Spectroscopy confirm that protein was incorporated into the APA micropores.

Section: Discussionsupporting

confidence: 74%

Protein Confinement in the Anodic Porous Alumina Microarray

Larosa¹,

Fiordoro²,

Nicolini³

et al. 2017

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“…LB nanotemplate method allows to improve the diffraction quality and radiation stability of protein crystals, as described in details in [42]. However, the RrA structure appears to be different from existing PDB asparaginases structures and models based on them.…”

Section: Resultsmentioning

confidence: 99%

LB Crystallization and Preliminary X-ray Diffraction Analysis of L-Asparaginase from Rhodospirillum rubrum

Pechkova¹,

Fiordoro²,

Sokolov³

et al. 2017

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Published by United Scientific Group AbstractProtein X-ray crystallography will remain the most powerful method to obtain the protein 3D atomic structures in foreseeable future. However, the production of the protein crystal as well as it quality (order, intensity of diffraction, radiation stability) remains the major problem. Many important proteins including those of life science interest and pharmaceutical industry impact are difficult to crystallize. The second major problem in protein crystallography is radiation damage of obtaining crystals which can only be partially overcome by existing methods. In the present work we use the protein LB nanotemplate crystallization methodgeneralized procedure for triggering of crystallization of any given protein, which allows to obtain radiation stable and high quality diffracting crystals for further X-ray analysis by synchrotron radiation. We apply LB nanotemplate method to crystallization of L-asparaginase from Rhodospirillum rubrum. This protein has potential application for combined chemical and enzymatic therapy of malignant blood disorders and therefore for new anticancer drug development. We also compare the diffraction quality of asparagines crystal obtained by classical method and LB nanotemplate and report preliminary X-ray diffraction characterization by synchrotron radiation.

“…The manuscript is carrying indeed the clear idea of stability of insulin crystallized by LangmuirBlodgett techniques, which is interesting and promising result which could have practical use in pharmaceutics. Limits and significance of this and previous in silico studies emerge from the recent most comprehensive publication in all aspects of LB nanotechnology [30]. …”

Section: Discussionmentioning

confidence: 99%

Langmuir-Blodgett Technology for Drugs Production and Delivery: Insights and Implications from an In Silico Study

Bragazzi¹,

Pechkova²,

Nicolini³

2017

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Insulin dynameomics here reported as a database pertaining to a series of Molecular Dynamics (MD) ran for Protein Data Bank (PDB) entries consisting in bioinformatics and molecular dynamics simulation of Langmuir-Blodgett and classical insulin with and without insulin receptor, appears to have profound implications for drug design and endocrinology. The slight differences in conformation and dynamics here reported may indeed explain why LB-insulin is more stable when binds to its receptor (lower free energy) and this could be useful when designing new drugs and pharmaceuticals.