Metabolic labeling of proteins with non-canonical amino acids (ncAAs) provides unique bioorthogonal chemical groups during de novo synthesis by taking advantage of both endogenous and heterologous protein synthesis machineries. Labeled proteins can then be selectively conjugated to fluorophores, affinity reagents, peptides, polymers, nanoparticles or surfaces for a wide variety of downstream applications in proteomics and biotechnology. In this review, we focus on techniques in which proteins are residue- and site-specifically labeled with ncAAs containing bioorthogonal handles. These ncAA-labeled proteins are: readily enriched from cells and tissues for identification via mass spectrometry-based proteomic analysis; selectively purified for downstream biotechnology applications; or labeled with fluorophores for in situ analysis. To facilitate the wider use of these techniques, we provide decision trees to help guide the design of future experiments. It is expected that the use of ncAA labeling will continue to expand into new application areas where spatial and temporal analysis of proteome dynamics and engineering new chemistries and new function into proteins are desired.

In breast cancer (BC), tissue stiffening via fibronectin (FN) and collagen accumulation is associated with advanced disease progression at both the primary tumor and metastatic sites. Here, we evaluate FN production in 15 BC cell lines, representing a variety of subtypes, phenotypes, metastatic potentials, and chemotherapeutic sensitivities. We demonstrate that intracellular and soluble FN is initially lost during tumorigenic transformation but is rescued in all lines with epithelial-mesenchymal plasticity (EMP). Importantly, we establish that no BC cell line was able to independently organize a robust FN matrix. Non-transformed mammary epithelial cells were also unable to deposit FN matrices unless transglutaminase 2, a FN crosslinking enzyme, was overexpressed. Instead, BC cells manipulated the FN matrix production of fibroblasts in a phenotypic-dependent manner. In addition, varied accumulation levels were seen depending if the fibroblasts were conditioned to model paracrine signaling or endocrine signaling of the metastatic niche. In the former, fibroblasts conditioned by BC cultures with high EMP resulted in the largest FN matrix accumulation. In contrast, mesenchymal BC cells produced extracellular vesicles (EV) that resulted in the highest levels of matrix formation by conditioned fibroblasts. Overall, we demonstrate a dynamic relationship between tumor and stromal cells within the tumor microenvironment, in which the levels and fibrillarization of FN in the extracellular matrix are modulated during the particular stages of disease progression.

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Purpose The purpose of this paper is to examine critically the different influences of open/closed innovation on employees’ performance. Design/methodology/approach The paper takes into account the different influence of open and closed innovation on the performance of employees. Some previous literature was reviewed and the quantitative method used a structured questionnaire to collect primary data from participants. Findings The results indicated that open innovation was more influential on the performance of employees than closed innovation. It also appeared that the case study organization uses the open innovation approach in the investments department to attract more clients and investments and thus increase its benefits. Practical implications The paper presents a critical account of differences in the influence on employees’ performance between open and closed innovation; it also takes into account the pros and cons of both aspects and their influence on the overall performance of the organization, putting into perspective the recommendations for the best approach to guarantee a good orientation within the internal environment of the organization. Finally, it helps in understanding the best approach to attract talent and creativity to the organization. Originality/value It is expected that the current research will offer guidance to organizations in Jordan to draw up plans for effective management of innovative approaches both internally and externally. It highlights the practice of open innovation and its role in attracting talent to the organization, to boost employees’ performance.

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