Rotational dynamics of actin

The rotational diffusion of actin was studied with the technique of time-resolved phosphorescence anisotropy using actin labeled at Cys-374 with erythrosin iodoacetamide. Immediately after the polymerization of actin was initiated, the correlation time increased sharply, passing through a maximum at 5 min and then declined to low values. F-Actin at equilibrium showed no anisotropy decay. The results were interpreted as indicating the initial formation of short mobile filaments which became increasingly immobile as elongation proceeded, leaving a decay which was dominated by shorter filaments. Some of these short filaments could have arisen by fragmentation of longer filaments. Eventually, the shorter filaments themselves became immobilized by entanglement within the gel matrix. The infinite-time anisotropy increased during polymerization, reflecting a smaller range of angular motion of the probe brought about by restricted torsional motion on the submicrosecond time scale. The results were compared with the length distribution of actin filaments revealed by electron microscopy [Kawamura, M., & Maruyama, K. (1970) J. Biochem. (Tokyo) 67, 437-457]. Polymerization in the presence of 1 microM cytochalasin B abolished the maximum in the correlation time profile and tended to prevent the immobilization of filaments by favoring shorter capped filaments which retained considerable rotational freedom. Addition of spectrin dimer to F-actin caused an increase in the time-invariant anisotropy. Subsequent additions of spectrin-binding proteins (erythrocyte bands 2.1 and 4.1) caused further increases in the anisotropy in a concentration-dependent manner, suggesting additional restriction of submicrosecond torsional motions. The results suggest that actin filaments within nonmuscle cells are rotationally immobile particularly if they are cross-linked by actin-binding proteins.     

Effect of ionic strength on the transfer of 1-pyrenemethyl-3 beta-hydroxy-22,23-bisnor-5-cholenate between bilayer vesicles containing phosphatidylserine.

The influence of ionic strength or the concentration of K+ ([K+]) of the aqueous phase on the spontaneous transfer of cholesterol between negatively charged bilayer vesicles composed of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylserine (DPPS) (1:1, mole:mole) was studied using a pyrene-labelled cholesterol analogue, 1-pyrenemethyl-3 beta-hydroxy-22,23-bisnor-5-cholenate (PMC), as the probe. The decrease in PMC excimer fluorescence was best fitted to a bi-exponential function. Increasing [K+] from 0.1 M to 0.3 M had little effect on the shorter half-time (1.4 +/- 0.2 min) but increased the longer half-time from 16.3 +/- 1.9 min to 26.7 +/- 2.1 min. Fluorescence quenching and titration of an interface-located fluorophore, 1-anilinonaphthalene-8-sulfonic acid (ANS) revealed an increase in interfacial hydrophobicity upon increasing in ionic strength. The physical state of the acyl chains was not affected by ionic strength as indicated by a constant PMC excimer:monomer fluorescence intensity ratio. However, an increase in enthalpy change of the lipid phase transition from 15.7 kJ/mol ([K+] = 0.1 M) to 21.3 kJ/mol ([K+] = 0.3 M), together with a slight increase in the transition temperature, implies that interactions between adjacent molecules in the charged lipid bilayer vesicles became stronger at higher ionic strength. Our results suggest that the van der Waals attraction between PMC and phospholipid molecules could be affected by conformation changes in the charged head group region brought about by changes of ionic strength in the aqueous phase, with consequent effects on the desorption of cholesterol from the bilayer surface.     

Migratory Disturbance Thresholds with Mule Deer and Energy Development

Fine-scale movement data has transformed our knowledge of ungulate migration ecology and now provides accurate, spatially explicit maps of migratory routes that can inform planning and management at local, state, and federal levels. Among the most challenging land use planning issues has been developing energy resources on public lands that overlap with important ungulate habitat, including the migratory routes of mule deer (Odocoileus hemionus). We generally know that less development is better for minimizing negative effects and maintaining habitat function, but we lack information on the amount of disturbance that animals can tolerate before reducing use of or abandoning migratory habitat. We used global positioning system data from 56 deer across 15 years to evaluate how surface disturbance from natural gas well pads and access roads in western Wyoming, USA, affected habitat selection of mule deer during migration and whether any disturbance threshold(s) existed beyond which use of migratory habitat declined. We used resource and step selection functions to examine disturbance thresholds at 3 different spatial scales. Overall, migratory use by mule deer declined as surface disturbance increased. Based on the weight of evidence from our 3 independent but complementary metrics, declines in migratory use related to surface disturbance were non-linear, where migratory use sharply declined when surface disturbance from energy development exceeded 3%. Disturbance thresholds may vary across regions, species, or migratory habitats (e.g., stopover sites). Such information can help with management and land use decisions related to mineral leasing and energy development that overlap with the migratory routes of ungulates. © 2020 The Wildlife Society.     

Planar cell polarity proteins differentially regulate extracellular matrix organization and assembly during zebrafish gastrulation

Zebrafish gastrulation cell movements occur in the context of dynamic changes in extracellular matrix (ECM) organization and require the concerted action of planar cell polarity (PCP) proteins that regulate cell elongation and mediolateral alignment. Data obtained using Xenopus laevis gastrulae have shown that integrin–fibronectin interactions underlie the formation of polarized cell protrusions necessary for PCP and have implicated PCP proteins themselves as regulators of ECM. By contrast, the relationship between establishment of PCP and ECM assembly/remodeling during zebrafish gastrulation is unclear. We previously showed that zebrafish embryos carrying a null mutation in the four-pass transmembrane PCP protein vang-like 2 (vangl2) exhibit increased matrix metalloproteinase activity and decreased immunolabeling of fibronectin. These data implicated for the first time a core PCP protein in the regulation of pericellular proteolysis of ECM substrates and raised the question of whether other zebrafish PCP proteins also impact ECM organization. In Drosophila melanogaster, the cytoplasmic PCP protein Prickle binds Van Gogh and regulates its function. Here we report that similar to vangl2, loss of zebrafish prickle1a decreases fibronectin protein levels in gastrula embryos. We further show that Prickle1a physically binds Vangl2 and regulates both the subcellular distribution and total protein level of Vangl2. These data suggest that the ability of Prickle1a to impact fibronectin organization is at least partly due to effects on Vangl2. In contrast to loss of either Vangl2 or Prickle1a function, we find that glypican4 (a Wnt co-receptor) and frizzled7 mutant gastrula embryos with disrupted non-canonical Wnt signaling exhibit the opposite phenotype, namely increased fibronectin assembly. Our data show that glypican4 mutants do not have decreased proteolysis of ECM substrates, but instead have increased cell surface cadherin protein expression and increased intercellular adhesion. These data indicate that Wnt/Glypican4/Frizzled signaling regulates ECM assembly through effects on cadherin-mediated cell cohesion. Together, our results demonstrate that zebrafish Vangl2/Prickle1a and non-canonical Wnt/Frizzled signaling have opposing effects on ECM organization underlying PCP and gastrulation cell movements.     

Spatial memory shapes migration and its benefits: evidence from a large herbivore

From fine‐scale foraging to broad‐scale migration, animal movement is shaped by the distribution of resources. There is mounting evidence, however, that learning and memory also guide movement. Although migratory mammals commonly track resource waves, how resource tracking and memory guide long‐distance migration has not been reconciled. We examined these hypotheses using movement data from four populations of migratory mule deer (n = 91). Spatial memory had an extraordinary influence on migration, affecting movement 2–28 times more strongly than tracking spring green‐up or autumn snow depth. Importantly, with only an ability to track resources, simulated deer were unable to recreate empirical migratory routes. In contrast, simulated deer with memory of empirical routes used those routes and obtained higher foraging benefits. For migratory terrestrial mammals, spatial memory provides knowledge of where seasonal ranges and migratory routes exist, whereas resource tracking determines when to beneficially move within those areas.     

Potent V2 vasopressin antagonists with structural changes at their C-terminals

A variety of structural changes were made in the C-terminals of four potent antidiuretic (V2) antagonists. The parent analogs were all derivatives of [1-(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid)]arginine-vasopressin, d(CH2)5AVP, namely d(CH2)5[D-Phe2,Ile4]AVP, d(CH2)5[D-Ile2,Ile4]AVP, d(CH2)5[D-Tyr(Et)2, Val4]AVP and d(CH2)5[D-Tyr(Et)2,Ile4]AVP. A number of amino acid amides were substituted for the C-terminal 9-glycinamide without reducing their V2-antagonistic potencies in rats. Many non-amino acid structures were also tolerated at the C-terminals of these antagonists and this end of these peptides can be prolonged without interfering with antagonistic potencies. Such altered V2-antagonists may be useful for the development of radioactive ligands, affinity labels and in affinity columns for studies on antidiuretic receptors. These C-terminal modifications also provide useful information for the further development of potent and specific V2-antagonists which can be valuable pharmacological tools and also promise to become useful clinically for the treatment of excessive water retention.     

Selective, high-affinity binding of ferric ions by glycine-extended gastrin(17)

Uptake of dietary iron is essential for replenishment of body stores. A role for the hormone gastrin in iron uptake as a chelator of ferric ions in the gastric lumen has been proposed previously [Baldwin, G. S. (1992) Med. Hypotheses 38, 70-74]. Here, spectroscopic evidence of selective, high-affinity binding of ferric ions to progastrin-derived peptides in aqueous solution at low pH is provided. The maximum at 281 nm in the absorption spectrum of glycine-extended gastrin(17) at pH 4.0 increased (2.07 +/- 0.30)-fold in the presence of > or =2 equiv of ferric ions. Titration of glycine-extended gastrin(17) with ferric ions under stoichiometric conditions indicated that the stoichiometry of binding was 2.00 +/- 0.28 mol of Fe(3+)/mol of peptide. Fluorescence quenching experiments yielded values for the stoichiometry and apparent dissociation constant of the ferric ion-glycine-extended gastrin(17) complex at pH 4.0 of 2.39 +/- 0.17 mol of Fe(3+)/mol and 0.62 +/- 0.19 microM, respectively. No interaction was detected with Co(2+), Cu(2+), Mn(2+), or Cr(3+). Electron paramagnetic resonance spectroscopy suggested that the iron ligands were either oxygen or sulfur atoms. Fluorescence quenching experiments with peptides derived from the glycine-extended gastrin(17) sequence indicated that one or more of the five glutamic acid residues were necessary for iron binding. The binding of ferric ions by glycine-extended gastrin(17) at low pH is consistent with a role for progastrin-derived peptides in iron uptake from the lumen of the gastrointestinal tract.