The three dimensional structure of Bovine Salivary Protein 30b (BSP30b) and its interaction with specific rumen bacteria

Mapping Intimacies ◽

10.1101/448167 ◽

2018 ◽

Author(s):

Heng Zhang ◽

Judith Burrows ◽

Graeme L. Card ◽

Graeme Attwood ◽

Thomas T. Wheeler ◽

...

Keyword(s):

Oleic Acid ◽

Binding Proteins ◽

Bacterial Species ◽

Fluorescent Microscopy ◽

Salivary Protein ◽

Lipid Binding ◽

Dimensional Structure ◽

Rumen Bacteria ◽

Abundant Protein

AbstractBovine Salivary Protein 30b (BSP30b) is a member of the tubular lipid-binding (TULIP) superfamily that includes the human bactericidal/permeability-increasing proteins (BPI), lipopolysaccharide binding proteins (LBP) and palate, lung, and nasal epithelium carcinoma-21 associated proteins (PLUNC). BSP30b is most closely related to the PLUNC family and is predominantly found in bovine saliva. There are four BSP30 isoforms (BSP30a-d) and collectively, they are the most abundant protein component of bovine saliva. The PLUNC family members are proposed to be lipid binding proteins, although in most cases their lipid ligands are unknown. Here, we present the X-ray crystal structure of BSP30b at 2.0 Å resolution. We used a double methionine mutant and Se-Met SAD phasing to solve the structure. The structure adopts a curved cylindrical form with a hydrophobic channel formed by an α/β wrap, which is consistent with the TULIP superfamily. The structure of BSP30b in complex with oleic acid is also presented where the ligand is accommodated within the hydrophobic channel. The electron density for oleic acid suggests that the ligand is only partially occupied in the binding site implying that oleic acid may not be the preferred ligand. GFP-tagged BSP30b binds to the surface of olive oil droplets, as observed under fluorescent microscopy, and acts as a surfactant consistent with its association with decreased susceptibility to bloat in cattle. Bacteria extracted directly from bovine rumen contents indicate that the GFP_BSP30b fusion protein binds to a small number of selected bacterial species in vivo. These results suggest that BSP30b may bind to bacterial lipids from specific species and that this abundant protein may have important biological roles via interacting with rumen bacteria during feeding and rumination.

Download Full-text

Surprising roles for phospholipid binding proteins revealed by high throughput geneticsThis paper is one of a selection of papers published in this special issue entitled “Second International Symposium on Recent Advances in Basic, Clinical, and Social Medicine” and has undergone the Journal's usual peer review process.

Biochemistry and Cell Biology ◽

10.1139/o09-171 ◽

2010 ◽

Vol 88 (4) ◽

pp. 565-574 ◽

Cited By ~ 8

Author(s):

Marissa A. LeBlanc ◽

Christopher R. McMaster

Keyword(s):

High Throughput ◽

Mammalian Cells ◽

Binding Proteins ◽

Essential Gene ◽

Peer Review Process ◽

Vesicular Transport ◽

Lipid Binding ◽

And Function ◽

Lipid Binding Proteins

Saccharomyces cerevisiae remains an ideal organism for studying the cell biological roles of lipids in vivo, as yeast has phospholipid metabolic pathways similar to mammalian cells, is easy and economical to manipulate, and is genetically tractable. The availability of isogenic strains containing specific genetic inactivation of each non-essential gene allowed for the development of a high-throughput method, called synthetic genetic analysis (SGA), to identify and describe precise pathways or functions associated with specific genes. This review describes the use of SGA to aid in elucidating the function of two lipid-binding proteins that regulate vesicular transport, Sec14 and Kes1. Sec14 was first identified as a phosphatidylcholine (PC) – phosphatidylinositol (PI) transfer protein required for viability, with reduced Sec14 function resulting in diminished vesicular transport out of the trans-Golgi. Although Sec14 is required for cell viability, inactivating the KES1 gene that encodes for a member of the oxysterol binding protein family in cells lacking Sec14 function results in restoration of vesicular transport and cell growth. SGA analysis identified a role for Kes1 and Sec14 in regulating the level and function of Golgi PI-4-phosphate (PI-4-P). SGA also determined that Sec14 not only regulates vesicular transport out of the trans-Golgi, but also transport from endosomes to the trans-Golgi. Comparing SGA screens in databases, coupled with genetic and cell biological analyses, further determined that the PI-4-P pool affected by Kes1 is generated by the PI 4-kinase Pik1. An important biological role for Sec14 and Kes1 revealed by SGA is coordinate regulation of the Pik1-generated Golgi PI-4-P pool that in turn is essential for vesicular transport into and out of the trans-Golgi.

Download Full-text

The Highly Abundant Protein Ag-lbp55 fromAscaridia galliRepresents a Novel Type of Lipid-binding Proteins

Journal of Biological Chemistry ◽

10.1074/jbc.m504474200 ◽

2005 ◽

Vol 280 (50) ◽

pp. 41429-41438 ◽

Cited By ~ 8

Author(s):

Rositsa Jordanova ◽

Georgi Radoslavov ◽

Peter Fischer ◽

Andrew Torda ◽

Friedrich Lottspeich ◽

...

Keyword(s):

Binding Proteins ◽

Lipid Binding ◽

Abundant Protein ◽

Lipid Binding Proteins

Download Full-text

Genetic manipulation of rumen bacteria: from potential to reality

Australian Journal of Agricultural Research ◽

10.1071/ar9960247 ◽

1996 ◽

Vol 47 (2) ◽

pp. 247 ◽

Cited By ~ 7

Author(s):

K Gregg ◽

G Allen ◽

C Beard

Keyword(s):

Molecular Genetics ◽

Genetic Manipulation ◽

Bacterial Species ◽

Genetic Material ◽

Rumen Bacteria ◽

Butyrivibrio Fibrisolvens ◽

Prevotella Ruminicola ◽

The Stability

The development of techniques for manipulating the molecular genetics of bacteria led naturally to suggestions for using this technology to alter rumen function. Despite early difficulties, methods are now available to insert new genetic material into several rumen bacterial species, including Butyrivibrio fibrisolvens, Prevotella ruminicola, and Ruminococcus albus. One strain of B. fibrisolvens has been modified to detoxify a naturally occurring poison that causes major losses of livestock in Australia, Africa, and Central America. The stability of that modified organism has been demonstrated by its recolonization of the rumen and retention of its altered genotype over 5 months in vivo. Many of the persistent doubts about rumen bacterial genetic manipulation and the viability of altered organisms in a competitive environment have been shown to be capable of resolution, and interest in this area of research may be revitalized by these results. Apart from the achievement of specific metabolic improvements, the technology now available will allow extensive characterization of the molecular genetics of rumen bacteria with a precision that was not previously possible.

Download Full-text

What Can Mushroom Proteins Teach Us about Lipid Rafts?

Membranes ◽

10.3390/membranes11040264 ◽

2021 ◽

Vol 11 (4) ◽

pp. 264

Author(s):

Maja Grundner ◽

Anastasija Panevska ◽

Kristina Sepčić ◽

Matej Skočaj

Keyword(s):

Lipid Rafts ◽

Lipid Raft ◽

Mammalian Cells ◽

Binding Proteins ◽

Protein Complexes ◽

Biological Membranes ◽

Indirect Evidence ◽

Lipid Binding ◽

Liquid Ordered

The lipid raft hypothesis emerged as a need to explain the lateral organization and behavior of lipids in the environment of biological membranes. The idea, that lipids segregate in biological membranes to form liquid-disordered and liquid-ordered states, was faced with a challenge: to show that lipid-ordered domains, enriched in sphingomyelin and cholesterol, actually exist in vivo. A great deal of indirect evidence and the use of lipid-binding probes supported this idea, but there was a lack of tools to demonstrate the existence of such domains in living cells. A whole new toolbox had to be invented to biochemically characterize lipid rafts and to define how they are involved in several cellular functions. A potential solution came from basic biochemical experiments in the late 1970s, showing that some mushroom extracts exert hemolytic activities. These activities were later assigned to aegerolysin-based sphingomyelin/cholesterol-specific cytolytic protein complexes. Recently, six sphingomyelin/cholesterol binding proteins from different mushrooms have been identified and have provided some insight into the nature of sphingomyelin/cholesterol-rich domains in living vertebrate cells. In this review, we dissect the accumulated knowledge and introduce the mushroom lipid raft binding proteins as molecules of choice to study the dynamics and origins of these liquid-ordered domains in mammalian cells.

Download Full-text

The three dimensional structure of Bovine Salivary Protein 30b (BSP30b) and its interaction with specific rumen bacteria

PLoS ONE ◽

10.1371/journal.pone.0206709 ◽

2019 ◽

Vol 14 (4) ◽

pp. e0206709

Author(s):

Heng Zhang ◽

Judith Burrows ◽

Graeme L. Card ◽

Graeme Attwood ◽

Tom T. Wheeler ◽

...

Keyword(s):

Three Dimensional ◽

Salivary Protein ◽

Dimensional Structure ◽

Rumen Bacteria ◽

Three Dimensional Structure

Download Full-text

Ubiquitin Activates Patatin-Like Phospholipases from Multiple Bacterial Species

Journal of Bacteriology ◽

10.1128/jb.02402-14 ◽

2014 ◽

Vol 197 (3) ◽

pp. 529-541 ◽

Cited By ~ 25

Author(s):

David M. Anderson ◽

Hiromi Sato ◽

Aaron T. Dirck ◽

Jimmy B. Feix ◽

Dara W. Frank

Keyword(s):

Bacterial Species ◽

Expression System ◽

Lipid Binding ◽

Amino Acid Sequences ◽

Cellular Damage ◽

Content Type ◽

Immunological Responses ◽

Binding Preference

Phospholipase A2enzymes are ubiquitously distributed throughout the prokaryotic and eukaryotic kingdoms and are utilized in a wide array of cellular processes and physiological and immunological responses. Several patatin-like phospholipase homologs of ExoU fromPseudomonas aeruginosawere selected on the premise that ubiquitin activation of this class of bacterial enzymes was a conserved process. We found that ubiquitin activated all phospholipases tested in bothin vitroandin vivoassays via a conserved serine-aspartate catalytic dyad. Ubiquitin chains versus monomeric ubiquitin were superior in inducing catalysis, and ubiquitin-like proteins failed to activate phospholipase activity. Toxicity studies in a prokaryotic dual-expression system grouped the enzymes into high- and low-toxicity classes. Toxicity measured in eukaryotic cells also suggested a two-tiered classification but was not predictive of the severity of cellular damage, suggesting that each enzyme may correspond to unique properties perhaps based on its specific biological function. Additional studies on lipid binding preference suggest that some enzymes in this family may be differentially sensitive to phosphatidyl-4,5-bisphosphate in terms of catalytic activation enhancement and binding affinity. Further analysis of the function and amino acid sequences of this enzyme family may lead to a useful approach to formulating a unifying model of how these phospholipases behave after delivery into the cytoplasmic compartment.

Download Full-text

Structural analysis of the photosynthetic membrane from Ectothiorhodospira halochloris

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010007727x ◽

1983 ◽

Vol 41 ◽

pp. 740-741

Author(s):

H. Engelhardt ◽

R. Guckenberger ◽

W. Baumeister

Keyword(s):

Lattice Structure ◽

Bacterial Species ◽

Hexagonal Lattice ◽

Reaction Centre ◽

Photosynthetic Membrane ◽

Rhodopseudomonas Viridis ◽

Photosynthetic Membranes ◽

Ectothiorhodospira Halochloris ◽

Main Components

Bacterial photosynthetic membranes contain, apart from lipids and electron transport components, reaction centre (RC) and light harvesting (LH) polypeptides as the main components. The RC-LH complexes in Rhodopseudomonas viridis membranes are known since quite seme time to form a hexagonal lattice structure in vivo; hence this membrane attracted the particular attention of electron microscopists. Contrary to previous claims in the literature we found, however, that 2-D periodically organized photosynthetic membranes are not a unique feature of Rhodopseudomonas viridis. At least five bacterial species, all bacteriophyll b - containing, possess membranes with the RC-LH complexes regularly arrayed. All these membranes appear to have a similar lattice structure and fine-morphology. The lattice spacings of the Ectothiorhodospira haloohloris, Ectothiorhodospira abdelmalekii and Rhodopseudomonas viridis membranes are close to 13 nm, those of Thiocapsa pfennigii and Rhodopseudomonas sulfoviridis are slightly smaller (∼12.5 nm).

Download Full-text

High resolution spot scan imaging of frozen, hydrated actin bundles with 400kV electrons

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122964 ◽

1992 ◽

Vol 50 (1) ◽

pp. 512-513

Author(s):

J. Jakana ◽

M.F. Schmid ◽

P. Matsudaira ◽

W. Chiu

Keyword(s):

High Resolution ◽

Binding Proteins ◽

Actin Binding ◽

Eukaryotic Cells ◽

Dimensional Structure ◽

Structural Basis ◽

Actin Bundle ◽

Actin Bundles ◽

3 Dimensional ◽

Macromolecular Assembly

Actin is a protein found in all eukaryotic cells. In its polymerized form, the cells use it for motility, cytokinesis and for cytoskeletal support. An example of this latter class is the actin bundle in the acrosomal process from the Limulus sperm. The different functions actin performs seem to arise from its interaction with the actin binding proteins. A 3-dimensional structure of this macromolecular assembly is essential to provide a structural basis for understanding this interaction in relationship to its development and functions.

Download Full-text

Production of Novel Bioactive Compounds by the Bacteria Associated with Some Marine Sponges of Gulf of Mannar and Palk Bay, Southeast Coast of India

International Journal of Emerging Research in Management and Technology ◽

10.23956/ijermt.v6i8.136 ◽

2018 ◽

Vol 6 (8) ◽

pp. 183

Author(s):

V. Ramadas ◽

G. Chandralega

Keyword(s):

Bioactive Compounds ◽

Bacterial Species ◽

Marine Sponges ◽

Marine Organisms ◽

Gulf Of Mannar ◽

Bacterial Symbionts ◽

Palk Bay ◽

Excellent Source

Sponges, exclusively are aquatic and mostly marine, are found from the deepest oceans to the edge of the sea. There are approximately 15,000 species of sponges in the world, of which, 150 occur in freshwater, but only about 17 are of commercial value. A total of 486 species of sponges have been identified in India. In the Gulf of Mannar and Palk Bay a maximum of 319 species of sponges have been recorded. It has been proved that marine organisms are excellent source of bioactive secondary metabolites and number of compounds of originated from marine organisms had been reported to possess in-vitro and in-vivo immuno stimulatory activity. Extracts from 20 sponge species were tested for bacterial symbionts and bioactive compounds were isolated from such associated bacterial species in the present study.

Download Full-text