A Receptor's Tale: An Eon in the Life of a Trypanosome Receptor

African trypanosomes have complex life cycles comprising at least ten developmental forms, variously adapted to different niches in their tsetse fly vector and their mammalian hosts. Unlike many other protozoan pathogens, they are always extracellular and have evolved intricate surface coats that allow them to obtain nutrients while also protecting them from the immune defenses of either insects or mammals. The acquisition of macromolecular nutrients requires receptors that function within the context of these surface coats. The best understood of these is the haptoglobin-hemoglobin receptor (HpHbR) of $\textit{Trypanosoma brucei}$, which is used by the mammalian bloodstream form of the parasite, allowing heme acquisition. However, in some primates it also provides an uptake route for trypanolytic factor-1, a mediator of innate immunity against trypanosome infection. Recent studies have shown that during the evolution of African trypanosome species the receptor has diversified in function from a hemoglobin receptor predominantly expressed in the tsetse fly to a haptoglobin-hemoglobin receptor predominantly expressed in the mammalian bloodstream. Structural and functional studies of homologous receptors from different trypanosome species have allowed us to propose an evolutionary history for how one receptor has adapted to different roles in different trypanosome species. They also highlight the challenges that a receptor faces in operating on the complex trypanosome surface and show how these challenges can be met.This has been funded by the Medical Research Council (Grant reference MR/L008246/1) and the Wellcome Trust (reference: 101020/Z/13/Z)

Evolutionary diversification of the trypanosome haptoglobin-haemoglobin receptor from an ancestral haemoglobin receptor.

The haptoglobin-haemoglobin receptor of the African trypanosome species, Trypanosoma brucei, is expressed when the parasite is in the bloodstream of the mammalian host, allowing it to acquire haem through the uptake of haptoglobin-haemoglobin complexes. Here we show that in Trypanosoma congolense this receptor is instead expressed in the epimastigote developmental stage that occurs in the tsetse fly, where it acts as a haemoglobin receptor. We also present the structure of the T. congolense receptor in complex with haemoglobin. This allows us to propose an evolutionary history for this receptor, charting the structural and cellular changes that took place as it adapted from a role in the insect to a new role in the mammalian host.Medical Research Counci

Structure and variability of the Denmark Strait Overflow: Model and observations

We report on a combined modeling and observational effort to understand the Denmark Strait Overflow (DSO). Four cruises over the course of 3 years mapped hydrographic properties and velocity fields with high spatial resolution. The observations reveal the mean path of the dense water, as well as the presence of strong barotropic flows, energetic variability, and strong bottom friction and entrainment. A regional sigma coordinate numerical model of interbasin exchange using realistic bottom topography and an overflow forced only by an upstream reservoir of dense fluid is compared with the observations and used to further investigate these processes. The model successfully reproduces the volume transport of dense water at the sill, as well as the 1000-m descent of the dense water in the first 200 km from the sill and the intense eddies generated at 1–3 day intervals. Hydraulic control of the mean flow is indicated by a region supercritical to long gravity waves in the dense layer located approximately 100 km downstream of the sill in both model and observations. In addition, despite the differences in surface forcing, both model and observations exhibit similar transitions from mostly barotropic flow at the sill to a bottom-trapped baroclinic flow downstream, indicating the dominant role of the overflow in determining the full water column dynamics

Hydrology in the Sea of Marmara during the last 23 ka : implications for timing of Black Sea connections and sapropel deposition

Author Posting. © American Geophysical Union, 2010. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Paleoceanography 25 (2010): PA1205, doi:10.1029/2009PA001735.Sediments deposited under lacustrine and marine conditions in the Sea of Marmara hold a Late Quaternary record for water exchange between the Black Sea and the Mediterranean Sea. Here we report a multiproxy data set based on oxygen and strontium isotope results obtained from carbonate shells, major and trace elements, and specific organic biomarker measurements, as well as a micropaleontological study from a 14C-dated sediment core retrieved from the Sea of Marmara. Pronounced changes occurred in δ18O and 87Sr/86Sr values at the fresh and marine water transition, providing additional information in relation to micropaleontological data. Organic biomarker concentrations documented the marine origin of the sapropelic layer while changes in n-alkane concentrations clearly indicated an enhanced contribution for organic matter of terrestrial origin before and after the event. When compared with the Black Sea record, the results suggest that the Black Sea was outflowing to the Sea of Marmara from the Last Glacial Maximum until the warmer Bølling-Allerød. The first marine incursion in the Sea of Marmara occurred at 14.7 cal ka B.P. However, salinification of the basin was gradual, indicating that Black Sea freshwaters were still contributing to the Marmara seawater budget. After the Younger Dryas (which is associated with a high input of organic matter of terrestrial origin) both basins were disconnected, resulting in a salinity increase in the Sea of Marmara. The deposition of organic-rich sapropel that followed was mainly related to enhanced primary productivity characterized by a reorganization of the phytoplankton population.We acknowledge support from INSU and the French Polar Institute IPEV

Structural analysis of haemoglobin binding by HpuA from the Neisseriaceae family

The Neisseriaceae family of bacteria causes a range of diseases including meningitis, septicaemia, gonorrhoea and endocarditis, and extracts haem from haemoglobin as an important iron source within the iron-limited environment of its human host. Herein we report crystal structures of apo- and haemoglobin-bound HpuA, an essential component of this haem import system. The interface involves long loops on the bacterial receptor that present hydrophobic side chains for packing against the surface of haemoglobin. Interestingly, our structural and biochemical analyses of Kingella denitrificans and Neisseria gonorrhoeae HpuA mutants, although validating the interactions observed in the crystal structure, show how Neisseriaceae have the fascinating ability to diversify functional sequences and yet retain the haemoglobin binding function. Our results present the first description of HpuA’s role in direct binding of haemoglobin

Structural insights into innate immunity against African trypanosomes

The haptoglobin-haemoglobin receptor (HpHbR) is expressed by the African try- panosome, T. brucei, whilst in the bloodstream of the mammalian host. This allows ac- quisition of haem, but also results in uptake of trypanolytic factor 1, a mediator of in- nate immunity against non-human African trypanosomes. Here, the structure of HpHbR in complex with its ligand, haptoglobin-haemoglobin (HpHb), is presented, revealing an elongated binding site along the membrane-distal half of the receptor. A &amp;Tilde;50° kink allows the simultaneous binding of two receptors to one dimeric HpHb, increasing the efficiency of ligand uptake whilst also increasing binding site exposure within the densely packed cell surface. The possibility of targeting this receptor with antibody-drug conjugates is ex- plored. The characterisation of the unexpected interaction between T. congolense HpHbR and its previously unknown ligand, haemoglobin, is also presented. This receptor is iden- tified as an epimastigote-specific protein expressed whilst the trypanosome occupies the mouthparts of the tsetse fly vector. An evolutionary pathway of the receptor is proposed, describing how the receptor has changed to adapt to a role as a bloodstream form-specific protein in T. brucei. Apolipoprotein L1 (ApoL1) is the pore-forming component of the trypanolytic factors. An expression and purification protocol for ApoL1 is presented here, and the functionality of the protein established. Initial attempts to characterise the pores and structure of ApoL1 are described.</p

A receptor's tale: An eon in the life of a trypanosome receptor

African trypanosomes have complex life cycles comprising at least ten developmental forms, variously adapted to different niches in their tsetse fly vector and their mammalian hosts. Unlike many other protozoan pathogens, they are always extracellular and have evolved intricate surface coats that allow them to obtain nutrients while also protecting them from the immune defenses of either insects or mammals. The acquisition of macromolecular nutrients requires receptors that function within the context of these surface coats. The best understood of these is the haptoglobin-hemoglobin receptor (HpHbR) of Trypanosoma brucei, which is used by the mammalian bloodstream form of the parasite, allowing heme acquisition. However, in some primates it also provides an uptake route for trypanolytic factor-1, a mediator of innate immunity against trypanosome infection. Recent studies have shown that during the evolution of African trypanosome species the receptor has diversified in function from a hemoglobin receptor predominantly expressed in the tsetse fly to a haptoglobin-hemoglobin receptor predominantly expressed in the mammalian bloodstream. Structural and functional studies of homologous receptors from different trypanosome species have allowed us to propose an evolutionary history for how one receptor has adapted to different roles in different trypanosome species. They also highlight the challenges that a receptor faces in operating on the complex trypanosome surface and show how these challenges can be met