Molecular physiology of the tensin brotherhood of integrin adaptor proteins.

Numerous proteins have been identified as constituents of the adhesome, the totality of molecular components in the supramolecular assemblies known as focal adhesions, fibrillar adhesions and other kinds of adhesive contact. The transmembrane receptor proteins called integrins are pivotal adhesome members, providing a physical link between the extracellular matrix (ECM) and the actin cytoskeleton. Tensins are ever more widely investigated intracellular adhesome constituents. Involved in cell attachment and migration, cytoskeleton reorganization, signal transduction and other processes relevant to cancer research, tensins have recently been linked to functional properties of deleted in liver cancer 1 (DLC1) and a mitogen-activated protein kinases (MAPK), to cell migration in breast cancer, and to metastasis suppression in the kidney. Tensins are close relatives of phosphatase homolog/tensin homolog (PTEN), an extensively studied tumor suppressor. Such findings are recasting the earlier vision of tensin (TNS) as an actin-filament (F-actin) capping protein in a different light. This critical review aims to summarize current knowledge on tensins and thus to highlight key points concerning the expression, structure, function, and evolution of the various members of the TNS brotherhood. Insight is sought by comparisons with homologous proteins. Some historical points are added for perspective.

Protein- and peptide-based electrospun nanofibers in medical biomaterials.

Electrospun fibers are being studied and developed because they hold considerable promise for realizing some advantages of nanostructured materials. The fibers can be made of biocompatible and biodegradable polymers. Electrospinning has therefore attracted interest in biotechnology and medicine, and there has been rapid growth in this area in recent years. This review presents an introduction to polymer nanofiber electrospinning, focusing on the use of natural proteins and synthetic peptides. We summarize key physical properties of protein-based and peptide-based nanofiber mats, survey biomedical applications of these materials, identify key challenges, and outline future prospects for development of the technology for tissue engineering, drug delivery, wound healing, and biosensors. FROM THE CLINICAL EDITOR: This review focuses on polymer nanofiber electrospinning using natural proteins and synthetic peptides. The authors describe key properties and applications of these materials, and outline future prospects for tissue engineering, drug delivery, wound healing, and biosensors based on these nanomats and nanofibers.

"In and out diffusion" hypothesis of exponential multilayer film buildup revisited.

A hypothesis concerning the exponential buildup of polyelectrolyte multilayer films prepared by layer-by-layer assembly has become widely accepted in the scientific community. This model was first introduced with experimental data in Langmuir. It was subsequently described in Proceedings of the National Academy of Sciences and extended and amended in papers in Langmuir and other journals. According to the "in and out diffusion" hypothesis, as it is called, or "common rule" of exponential multilayer film buildup, as it is widely regarded, "a diffusion-based buildup mechanism ... explains most of the exponential-like growth process of polyelectrolyte multilayers reported in the literature." The present work offers an alternative viewpoint to specific elements of the hypothesis and the model as a whole.

Present and future prospects for polypeptide multilayer nanofilms in biotechnology and medicine.

Polypeptide multilayer nanofilms are a promising nanotechnology for commercial product development because the processes used to prepare them are simple, flexible, reliable, automatable, and scalable. Moreover, these materials can display a remarkable diversity of physical, chemical, and biological properties. Furthermore, the constituents of these nanofilms, in most cases the nanofilms themselves, and the fabrication process are environmentally benign. Nanofilm structure and function can be tailored to address two Grand Challenges of the US National Nanotechnology Initiative.

Reversibility of structural changes of polypeptides in multilayer nanofilms.

Structural properties of different polypeptide multilayer nanofilms fabricated at neutral pH have been analyzed by UV spectroscopy, circular dichroism spectroscopy (CD), and Fourier-transform infrared spectroscopy (FTIR). The various peptides studied exhibit a strong tendency to adopt a beta sheet conformation in the films. Changes in film structure on dehydration are completely reversed on rewetting. The time scale of reversibility is, however, substantially shorter for the polymer backbone than the side chains, as in protein folding.

Stimulated release of small molecules from polyelectrolyte multilayer nanocoatings.

Free thiol-containing polyelectrolytes serve simultaneously as a material for self-assembly of a multilayer nanocoating and as a carrier of small molecules for release from the coating in response to an environmental cue.

Polypeptide multilayer nanofilm artificial red blood cells.

Reliable encapsulation of hemoglobin (Hb) within polypeptide multilayer nanofilms has been achieved by a template-based approach, and protein functionality has been demonstrated postencapsulation. The method is general in scope and could be useful for many other encapsulants. Met-Hb was adsorbed onto 5 microm-diameter CaCO3 microparticles, and the Hb-coated particles were encapsulated within a multilayer nanofilm of poly(L-glutamic acid) (PLGA) and poly(L-lysine) (PLL) by layer-by-layer assembly. The CaCO3 templates were then dissolved within the PLGA/PLL nanofilms by addition of ethylenediaminetetraacetic acid. Encapsulation of Hb was proved by fluorescence microscopy, the pH-dependence of retention of Hb was determined by visible wavelength absorbance, and conversion of the encapsulated met-Hb to deoxy-Hb and oxy-Hb was demonstrated by spectroscopic analysis of the Soret absorption peak under various conditions. It thus has been shown that control of Hb oxygenation within polypeptide multilayer nanofilm artificial cells is possible, and that Hb thus encapsulated can bind, release, and subsequently rebind molecular oxygen. This work therefore represents an advance in the development of polypeptide multilayer film artificial red blood cells.

Controlled loading and release of a model drug from polypeptide multilayer nanofilms.

A major concern of medicine today is the sustained release of therapeutic compounds. Delivery vehicles for such compounds must be biocompatible. Ideally, loading a drug into the delivery vehicle will be a simple process, and vehicle properties will allow control over the drug release profile under desired conditions. Here, polypeptide multilayer nanofilms have been prepared by electrostatic layer-by-layer self-assembly to study the post-fabrication loading and release of a model therapeutic, methylene blue (MB). Drug loading and release have been characterized by optical spectroscopy for different peptide designs at different pH values, and film surface morphology has been characterized by atomic force microscopy (AFM). Differences in peptide structure have been found to influence MB loading and release under otherwise fixed conditions. Release is also influenced by pH, salt concentration, and number of "capping" layers. Although more research will be needed to exhaust the potential of polypeptide multilayer films, present results would suggest that the technology holds considerable promise for applications in medicine.

From bench to bedside: successful translational nanomedicine: highlights of the Third Annual Meeting of the American Academy of Nanomedicine.

The Third Annual Meeting of the American Academy of Nanomedicine (AANM) was held at the University of California San Diego, in San Diego, California during September 7-8, 2007. The meeting was focused on successful translational nanomedicine: from bench to bedside. There were four keynote lectures and eight scientific symposiums in this meeting. The researchers and investigators reported the results and process of current nanomedicine research and approaches to clinical applications. The meeting provided exciting information for nanomedicine clinical-related researches and strategy for further development of nanomedicine research which will be benefits to clinical practice.

High-capacity functional protein encapsulation in nanoengineered polypeptide microcapsules.

Addition of polyethylene glycol to aqueous assembly solutions of oppositely charged polypeptides enables high-capacity "loading" of functional protein in biocompatible microcapsules by template-supported layer-by-layer nanoassembly.

Structural stability of polypeptide nanofilms under extreme conditions.

Self-assembly of designed peptides is a promising area of biomaterials research and development. Here, polypeptide nanofilms have been prepared by electrostatic layer-by-layer self-assembly (LBL) of cysteine (Cys)-containing 32mers designed to be oppositely charged at neutral pH, and structural stability of the films has been probed by subjecting them to various extreme physical and chemical conditions. The results suggest that although electrostatic attraction plays a key role in strengthening polypeptide films, stability is inversely related to absolute net charge of the supramolecular complex. This behavior is similar to the typical behavior of small globular proteins. Film structure is very stable in organic solvent and, when dehydrated, at extreme temperatures. Such stability is in marked contrast to the behavior of proteins, which tend to denature under comparable conditions. Similar to proteins, peptide nanofilms cross-linked by disulfide (S-S) bonds are considerably stronger than films stabilized by electrostatic, van der Waals, or hydrophobic interactions alone. This effect is particularly evident at extremes of pH and at elevated temperature when the film is hydrated. These results, the great variety of possible peptide structures, the inherent biocompatibility of l-amino acids, and current applications of thin films in commercial products together suggest that polypeptide films are promising for the development of new or enhanced products in food technology, drug delivery and medical device coatings, and biomaterials.

Fine tuning of physical properties of designed polypeptide multilayer films by control of pH.

Adjustment of pH can alter the ensemble of three-dimensional structures of a polypeptide in solution by changing the distribution of charge and Coulombic interactions. The role of pH in layer-by-layer self-assembly (LbL) of designed 32mer peptides containing the amino acid cysteine has been investigated using a combination of physical methods. Results show that pH can have a substantial influence on the mass of adsorbed peptide, surface roughness, and film density over a range of 1.5 pH units. Peptide film thickness depends on the number of layers, as with "conventional" polyelectrolytes. Film density and morphology, however, vary more with pH than does thickness, translating into a change in density on the order of 70% over the pH range 7.4-8.9. Results of this work provide insight on the physical basis of LbL and suggest that peptides are a promising class of polyelectrolytes for the creation of designer thin films for applications in biotechnology and other areas.

Physics of polypeptide multilayer films.

Polypeptide multilayer films are promising for the development of coatings for implant devices, biosensors, and artificial cells. This paper discusses aspects of the physics of these films. Three sub-topics in the physics of peptide adsorption in multilayer film assembly covered here are peptide structure at the film/solid support interface, adsorbed layer thickness, and dynamics of peptide adsorption. A synopsis of work in these areas is preceded by an introduction to the subject and a review of some aspects of polymer theory.

Protein-inspired multilayer nanofilms: science, technology and medicine.

The field of polypeptide multilayer nanofilm research flourishes where study of protein structure and function shares a border with development of polyelectrolyte multilayers. The soil is fertile for creative input and promises a harvest of interesting results: the structure of a film can be predetermined on a layer-by-layer (LBL) basis, a huge variety of polypeptide sequences can be realized in large quantities by modern methods of synthesis, and the fabrication process is environmentally benign. In electrostatic LBL assembly, multilayer film assembly is driven primarily by coulombic interactions, but hydrophobic interactions and hydrogen bonds also contribute to film formation and stability, the amount depending on polypeptide design. Most peptides suitable for LBL assembly form films with a large percentage of beta-sheet at neutral pH; it would appear that beta-sheet is favored over alpha-helix in this context by the contribution to entropy of the number of ways of forming a beta-sheet from a single polypeptide chain. Film thickness and roughness depend rather substantially on amino acid composition. Promising applications of the polypeptide multilayer film platform technology include coatings for medical implant devices, scaffolds for tissue engineering, coatings for targeted drug delivery, artificial cells for oxygen therapeutics, and artificial viruses for immunization. In each case peptide structure is tailored to the application. Here we summarize recent results of experimental studies and computational work from our laboratory, showing how the study of protein structure has inspired the design of polypeptide films and pointing out new opportunities for technology development. This work also provides a brief introduction to polypeptide structure and multilayer films.

New technology and clinical applications of nanomedicine: highlights of the second annual meeting of the American Academy of Nanomedicine (Part I).

The Second Annual Meeting of the American Academy of Nanomedicine (AANM) was held at the National Academy of Science Building in Washington, DC, September 9-10, 2006. The program included two Nobel Prize Laureate Lectures, two Keynote Lectures, and 123 invited outstanding State-in-Art lectures presenting in 23 special concurrent symposia. In addition, there were 22 poster presentations in the meeting addressing different areas in nanomedicine research. All of the presenters at the meeting are outstanding investigators and researchers in the field. The Second Annual Meeting of the AANM was a great success. The meeting provides investigators from different world areas a forum and an opportunity for discussion. We believe that nanomedicine research will develop rapidly in the future. The AANM invites basic and clinical researchers from the world to join this exciting research.

FDTD simulation of exposure of biological material to electromagnetic nanopulses.

Ultra-wideband (UWB) electromagnetic pulses of nanosecond duration, or nanopulses, are of considerable interest to the communications industry and are being explored for various applications in biotechnology and medicine. The propagation of a nanopulse through biological matter has been computed using the finite difference-time domain (FDTD) method. The approach required the reparametrization of existing Cole-Cole model-based descriptions of dielectric properties of biological matter in terms of the Debye model without loss of accuracy. Several tissue types have been considered. Results show that the electromagnetic field inside biological tissue depends on incident pulse rise time and width. Rise time dominates pulse behaviour inside tissue as conductivity increases. It has also been found that the amount of energy deposited by 20 kV m(-1) nanopulses is insufficient to change the temperature of the exposed material for pulse repetition rates of 1 MHz or less, consistent with recent experimental results.

Nanoscale biomimetics: fabrication and optimization of stability of peptide-based thin films.

The method of thin film preparation known as layer-by-layer assembly is of growing interest for current and envisioned developments in bionanotechnology. Here, cysteine-containing 32mer peptides have been designed, synthesized, purified, and used to prepare polypeptide films. A range of methods-quartz crystal microbalance, Fourier transform infrared spectroscopy, circular dichroism spectroscopy, and high-performance liquid chromatography-have been used to probe the effect of ionic strength and polymer secondary structure in solution on peptide self-assembly, and on secondary structure formation and disulfide bond cross-linking in the multilayer film. The amount of designed peptide deposited per adsorption step of film fabrication increased with increasing ionic strength, as with conventional polyelectrolytes. Secondary structure content changed from random coil to beta sheet on incorporation of peptides into a film. "Peptide-inherent" cross-linking by disulfide bond formation increased film stability at acidic pH. Conditions for disulfide stabilization have been optimized. The results contribute to exploration of the physical basis of peptide self-assembly broaden the scope of applications of layer-by-layer assembly, particularly where biocompatibility and stability are key design concerns, and provide a basis for mass production of custom polypeptide thin films of high stability, even in harsh environments.

Superdomains in the protein structure hierarchy: The case of PTP-C2.

Superdomain is uniquely defined in this work as a conserved combination of different globular domains in different proteins. The amino acid sequences of 25 structurally and functionally diverse proteins from fungi, plants, and animals have been analyzed in a test of the superdomain hypothesis. Each of the proteins contains a protein tyrosine phosphatase (PTP) domain followed by a C2 domain. Four novel conserved sequence motifs have been identified, one in the PTP domain and three in the C2 domain. All contribute to the PTP-C2 domain interface in PTEN, a tumor suppressor, and all are more conserved than the PTP signature motif, HCX3 (K/R)XR, in the 25 sequences. We show that PTP-C2 was formed prior to the fungi, plant, and animal kingdom divergence. A superdomain as defined here does not fit the usual protein structure classification system. The demonstrated existence of one superdomain suggests the existence of others.

Branched polymer models and the mechanism of multilayer film buildup.

The "in and out diffusion" hypothesis does not provide a conclusive explanation of the buildup displayed by some polyelectrolyte multilayer film systems. Here, we report initial tests of an alternative hypothesis, on which the completion of each adsorption cycle results in an increase in the number of polymer binding sites on the film surface. Polycationic dendrimeric peptides, which can potentially bind several oppositely-charged peptides each, have been designed, synthesized and utilized in comparative film buildup experiments. Material deposited, internal film structure and film surface morphology have been studied by ultraviolet spectroscopy (UVS), circular dichroism spectroscopy (CD), quartz crystal microbalance (QCM) and atomic force microscopy (AFM). Polycations tended to contribute more to film buildup than did polyanions on quartz but not on gold. Increasing the number of branches in the dendrimeric peptides from 4 to 8 reproducibly resulted in an increase in the film growth rate on quartz but not on gold. Peptide backbones tended to adopt a ß-strand conformation on incorporation into a film. Thicker films had a greater surface roughness than thin films. The data are consistent with film buildup models in which the average number of polymer binding sites will increase with each successive adsorption cycle in the range where exponential growth is displayed.