Copper(II) binding by the earliest vertebrate gonadotropin‐releasing hormone, the type II isoform, suggests an ancient role for the metal

In vertebrate reproductive biology copper can influence peptide and protein function both in the pituitary and in the gonads. In the pituitary, copper binds to the key reproductive peptides gonadotropin‐releasing hormone I (GnRH‐I) and neurokinin B, to modify their structure and function, and in the male gonads, copper plays a role in testosterone production, sperm morphology and, thus, fertility. In addition to GnRH‐I, most vertebrates express a second isoform, GnRH‐II. GnRH‐II can promote testosterone release in some species and has other non‐reproductive roles. The primary sequence of GnRH‐II has remained largely invariant over millennia, and it is considered the ancestral GnRH peptide in vertebrates. In this work, we use a range of spectroscopic techniques to show that, like GnRH‐I, GnRH‐II can bind copper. Phylogenetic analysis shows that the proposed copper‐binding ligands are retained in GnRH‐II peptides from all vertebrates, suggesting that copper‐binding is an ancient feature of GnRH peptides

Essential role of histidine for rapid copper(II)-mediated disassembly of neurokinin B amyloid

Neurokinin B is a tachykinin peptide involved in a diverse range of neuronal functions. It rapidly forms an amyloid, which is considered physiologically important for efficient packing into dense core secretory vesicles within hypothalamic neurons. Disassembly of the amyloid is thought to require the presence of copper ions, which interact with histidine at the third position in the peptide sequence. However, it is unclear how the histidine is involved in the amyloid structure and why copper coordination can trigger disassembly. In this work, we demonstrate that histidine contributes to the amyloid structure via π-stacking interactions with nearby phenylalanine residues. The ability of neurokinin B to form an amyloid is dependent on any aromatic residue at the third position in the sequence; however, only the presence of histidine leads to both amyloid formation and rapid copper-induced disassembly

The emerging role of astrocytes in metal homeostasis in the brain

Disruption to metal homeostasis is a clear and common feature of several debilitating neurodegenerative disorders. Over the last couple of decades significant advances have occurred in our understanding of biological metal homeostasis, particularly for the three most abundant metals, iron, zinc and copper. In contrast, despite having a requirement for these metal ions it is not clear how the brain controls and mediates uptake and distribution. However, it is emerging that a key component in metal homeostasis is glial cells, particularly astrocytes, and particularly those that interact with blood vessels and neurons. Astrocytes are known to help the regulation of the biochemical environment of the brain and are likely to assist in metal homeostasis. The use of metals in the brain (particularly copper and iron), the neurologic diseases that feature disrupted metal homeostasis and how astrocytes play a not insignificant role in metal regulation are the focus of this review

Metal-minded : use, control and metallic interplay of copper in the brain

Copper is the third most abundant transition metal in biological systems, and is a cofactor for a variety of proteins and enzymes. The ability of copper to cycle between oxidation states under physiological conditions is one reason for its versatility throughout many aspects of biology. The way in which cells ensure copper reaches its targets, yet limit redox activity, has become an area of widespread study over the past decade or so. It is now established that a variety of mechanisms are in place to successfully use copper whilst preventing uncontrolled, damaging redox activity. The study of metals in the central nervous system, encompassing the brain and spinal cord and associated nerves, is a burgeoning area of study, termed ‘metalloneurochemistry’, and the specialized nature of the brain suggests it is likely to have unique uses and homeostatic mechanisms. Despite its known involvement in diseases such as Menkes and Prion disorders, there is still much to be learnt about the physiological function of copper and the homeostatic mechanisms that control it under normal conditions in the brain. In this chapter we will outline recent work that investigates how copper is used in the brain, the control of metal concentrations and the interplay between copper and other metal ions, notably iron

Nitroxide spin-labelling and its role in elucidating cuproprotein structure and function

Copper is one of the most abundant biological metals, and its chemical properties mean that organisms need sophisticated and multilayer mechanisms in place to maintain homoeostasis and avoid deleterious effects. Studying copper proteins requires multiple techniques, but electron paramagnetic resonance (EPR) plays a key role in understanding Cu(II) sites in proteins. When spin-labels such as aminoxyl radicals (commonly referred to as nitroxides) are introduced, then EPR becomes a powerful technique to monitor not only the coordination environment, but also to obtain structural information that is often not readily available from other techniques. This information can contribute to explaining how cuproproteins fold and misfold. The theory and practice of EPR can be daunting to the non-expert; therefore, in this mini review, we explore how nitroxide spin-labelling can be used to help the inorganic biochemist gain greater understanding of cuproprotein structure and function in vitro and how EPR imaging may help improve understanding of copper homoeostasis in vivo

Neurokinin B and serum albumin limit copper binding to mammalian gonadotropin releasing hormone

Gonadotropin releasing hormone (GnRH) triggers secretion of luteinizing hormone and follicle stimulating hormone from gonadotropic cells in the anterior pituitary gland. GnRH is able to bind copper, and both invitro and invivo studies have suggested that the copper-GnRH complex is more potent at triggering gonadotropin release than GnRH alone. However, it remains unclear whether copper-GnRH is the active species invivo. To explore this we have estimated the GnRH-copper affinity and have examined whether GnRH remains copper-bound in the presence of serum albumin and the neuropeptide neurokinin B, both copper-binding proteins that GnRH will encounter invivo. We show that GnRH has a copper dissociation constant of 0.9 * 10-9 M, however serum albumin and neurokinin B can extract metal from the copper-GnRH complex. It is therefore unlikely that a copper-GnRH complex will survive transit through the pituitary portal circulation and that any effect of copper must occur outside the bloodstream in the absence of neurokinin B

The magic of partnering with students to create transdisciplinary STEM curricula

CONTEXT: Co-creation of curriculum with a range of different partners is an increasing practice of learning and teaching in the higher education sector. In research, partnership models for collaboration with a range of stakeholders, notably industry, are well established and practiced, but fewer models exist that show how to partner in teaching. The growing success stories of curriculum co-creation with student partners show that partnering with students is a practical and a sustainable way to create engaging curricula. A benefit is that student outcomes and desires often align with institutional aims such as producing capable and confident graduates that can work in any industry beyond geographical and disciplinary constraints. This contrasts with industry partners who are often interested in job ready graduates for the specific industry or sector. In this study we are presenting our learnings from partnering with students in designing transdisciplinary curricula. PURPOSE: The aim was to design minors that attract and encourage students from non-STEM backgrounds to use their elective space to study curriculum that helps them to develop STEM capabilities. This aim is in line with training job ready graduates and strategies to address National Priorities and Industry Linkage Fund (NPILF). The minors were designed in consultation and collaboration with industry partners and academics from different disciplines. APPROACH: Two minors were designed by choosing subjects from four different disciplines and a capstone project was added to create a hands-on learning opportunity for students. Subjects were chosen from four different schools to ensure the trans-disciplinarity aspect of the design. The most important criterion for subjects included in a minor was the lack of restrictive prerequisites to ensure that all students could enrol. OUTCOMES: Collaboration with Industry allowed us to implement development of STEM skills or capabilities as the learning outcome, but collaboration with students was instrumental in designing curricula that was attractive to students and relevant to the future generation of graduates. Therefore, students were embedded in the process of design of these minors all the way from ideation to marketing the curricula. Minors were built around two topics of health and sustainability that a) students found interesting and b) where industry indicated there were current and future job opportunities. The curriculum design was fine-tuned following several discussions with student partners and surveying a larger group of students. In consultation with subject coordinators the student partners critically evaluated the assessments in each of the subjects to ensure that students from diverse disciplinary backgrounds would be able to complete the subject. The minors were presented to a larger group of students before presenting them to various academic committees for approval. In addition to developing traditional resources such as the handbook entry, marketing videos were produced by student partners to communicate the new curricula to incoming students. “Innovating For Humans“ and “Eco-Socially Conscious Design & Manufacturing” were offered in the first academic session of 2022. CONCLUSIONS: Designing trans- multi or interdisciplinary, curricula is not just bringing subjects with no prerequisites from different disciplines around a core topic. It is vital to ensure that the subjects are indeed linked by a common theme, but that learning outcomes are scaffolded and achievable and that the subjects can be completed successfully by students from all disciplines. We soon discovered that learning guides were not the most reliable source of information and going through the learning activities and assessments with student partners and unit coordinators was essential to identify the hidden assumed knowledge and disciplinary focused skills. Subsequently, learning activities and assessment of some subjects were revised and the unit coordinators designed extra resources and supports to assist students from other disciplines. This process also benefits students from within the discipline. As STEM educators we did not anticipate that the biggest challenge of designing transdisciplinary curricula in STEM was to get the right level of STEM content to achieve the intended learning outcomes while keeping them attractive to students from other disciplines that were raised in an education system that presented STEM as a “difficult subject”. The outcome and process of this curriculum development work would have been very different without collaboration with student partners from different disciplines. Working with students was a great learning experience that can be described as designing a product for the end users with them and eliminating assumptions and predictions. Although this work was on designing interdisciplinary curricula, we believe this model is an efficient strategy that can be applied in designing of engaging discipline focused curriculum

[In Press] Endocytic recycling prevents copper accumulation in astrocytoma cells stimulated with copper-bound neurokinin B

Neurokinin B (NKB) is a key neuropeptide in reproductive endocrinology where it contributes to the generation of pulses of gonadotropin-releasing hormone. NKB is a copper-binding peptide; in the absence of metal NKB rapidly adopts an amyloid structure, but copper binding inhibits amyloid formation and generates a structure that can activate the neurokinin 3 receptor. The fate of copper once it binds NKB and activates the neurokinin 3 receptor is not understood, but endocytosis of NKB occurs even when the peptide is coordinated to copper. Using astrocytoma cells that express endogenous neurokinin 3 receptor, this work shows that endocytosis of apo- and copper-bound NKB occurs in concert with the receptor via a trafficking pathway that includes the early endosome. When cells are stimulated with copper-bound NKB the cellular copper concentration does not significantly increase, however when the cells are pre-treated with the recycling inhibitor, brefeldin A, they are capable of accumulating copper. This data shows that copper-bound NKB can activate the neurokinin 3 receptor then endocytosis abstracts metal, peptide and receptor from the cell surface. The cell does not accumulate the copper but instead it enters recycling pathways that ultimately leads to metal release from the cell. The work reveals a novel receptor-mediated copper trafficking pathway that retains metal in membrane bound organelles until it is exported from the cell

Endocytosis of G protein-coupled receptors and their ligands : is there a role in metal trafficking?

The prevalence of metal dysregulation in many neurodegenerative and neurocognitive disorders has compelled many studying such diseases to investigate the mechanisms underlying metal regulation in the central nervous system. Metal homoeostasis is often complex, with sophisticated, multilayered pathways in operation. G protein-coupled receptors are omnipresent on cell membranes and have intriguing mechanisms of endocytosis and trafficking that may be useful in metal homoeostasis. Indeed, many receptors and/or their cognate ligands are able to bind metals, and in many cases metals are considered to have neuromodulatory roles as a result of receptor binding. In this mini-review, we outline the structural and functional aspects of G protein-coupled receptors with a focus on the mechanisms leading to endocytosis and cellular trafficking. We further highlight how this may help in the trafficking of metal ions, notably copper

Development of a WIL rubric to facilitate identification and mapping of WIL activities in science courses

Identification of WIL activities occurring within units is necessary to allow accurate mapping across degrees and to provide assurance that WIL-related learning outcomes are being attained. Science degrees are challenging because some WIL activities are so highly visible (e.g. field work, laboratory practicals) that other valid activities are not recognised. These other activities, which we term ‘hidden WIL’, are often a rich source of WIL that can be directly related to employability, but are not articulated to students and sometimes not appreciated by the unit coordinator. Several WIL activity rubrics exist, with the recent Edwards review into WIL in Science providing a clear framework linking ‘learning outcomes’ to activities. We chose to use this framework to identify all WIL activities occurring across a diverse suite of science degrees. Using a questionnaire we interviewed the unit coordinators of 81 science units and identified both overt (clearly articulated to students) and hidden WIL in the units. We then mapped these activities to reconstruct the WIL content of complete degrees. We found that WIL activities were generally focused on a career as a research academic, and that hidden WIL was more prevalent in early years than in the final year, suggesting that academic staff find employability skills only become important near graduation. After using the Edwards framework we suggest refinements and additions and present a WIL activity rubric that will have broad applicability across disciplines