Complexity of Gas Vesicle Biogenesis in Halobacterium sp. Strain NRC-1: Identification of Five New Proteins

ABSTRACT The genome of Halobacterium sp. strain NRC-1 contains a large gene cluster, gvpMLKJIHGFEDACNO, that is both necessary and sufficient for the production of buoyant gas-filled vesicles. Due to the resistance of gas vesicles to solubilization, only the major gas vesicle protein GvpA and a single minor protein, GvpC, were previously detected. Here, we used immunoblotting analysis to probe for the presence of gas vesicle proteins corresponding to five additional gvp gene products. Polyclonal antisera were raised in rabbits against LacZ-GvpF, -GvpJ, and -GvpM fusion proteins and against synthetic 15-amino-acid peptides from GvpG and -L. Immunoblotting analysis was performed on cell lysates of wild-type Halobacterium sp. strain NRC-1, gas vesicle-deficient mutants, and purified gas vesicles, after purification of LacZ fusion antibodies on protein A and β-galactosidase affinity columns. Our results show the presence of five new gas vesicle proteins (GvpF, GvpG, GvpJ, GvpL, and GvpM), bringing the total number of proteins identified in the organelles to seven. Two of the new gas vesicle proteins are similar to GvpA (GvpJ and GvpM), and two proteins contain predicted coiled-coil domains (GvpF and GvpL). GvpL exhibited a multiplet ladder on sodium dodecyl sulfate-polyacrylamide gels indicative of oligomerization and self-assembly. We discuss the possible functions of the newly discovered gas vesicle proteins in biogenesis of these unique prokaryotic flotation organelles.

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Heterologous expression of cyanobacterial gas vesicle proteins in Saccharomyces cerevisiae

Biotechnology Journal ◽

10.1002/biot.202100059 ◽

2021 ◽

pp. 2100059

Author(s):

Harin Jung ◽

Hua Ling ◽

Yong Quan Tan ◽

Nam‐Hai Chua ◽

Wen Shan Yew ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Heterologous Expression ◽

Gas Vesicle Proteins ◽

Vesicle Proteins

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Solubilization and immunoprecipitation of 125I-labelled antigens from Plasmodium knowlesi schizont-infected erythrocytes using non-ionic, anionic and zwitterionic detergents

Parasitology ◽

10.1017/s0031182000054317 ◽

1984 ◽

Vol 88 (1) ◽

pp. 27-36 ◽

Cited By ~ 6

Author(s):

R. J. Howard ◽

J. W. Barnwell

Keyword(s):

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Plasmodium Knowlesi ◽

Polyacrylamide Gel Electrophoresis ◽

Protein A ◽

Sodium Dodecyl ◽

Ionic Detergents ◽

Class A ◽

Anionic Detergents ◽

Triton X

SUMMARYPlasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200000 and 180000 were detected only after extraction with deoxycholate or SDS.

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A New Approach to Fluorescent Chemosensor of Cu2+ Ion

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.513-517.65 ◽

2014 ◽

Vol 513-517 ◽

pp. 65-69

Author(s):

Xiao Jun Hu ◽

Xin Yan Hu ◽

Zhi Zhang

Keyword(s):

Aqueous Solution ◽

Electron Transfer ◽

Self Assembly ◽

Ph Value ◽

Sodium Dodecyl ◽

Fluorescence Emission ◽

Fluorescent Chemosensor ◽

Dynamic Quenching ◽

Transfer Effects ◽

New Approach

According to the principle of dynamic quenching a new ON-OFF fluorescent chemosensor for Cu2+ions was designed, this chemosensor was composed of p-tert-butylthiacalix [arene (TCA),sodium dodecyl sulfate (SDS) and perylene through the form of self-assembly in aqueous solution. Addition of Cu2+ions could result in a quenching of the fluorescence emission of perylene inside micelles, which due to intramicellar complex-fluorophore electron-transfer or energy-transfer effects induced by the complexation of TCA with the Cu2+ions.The experimental results indicated that: Under the condition of TCA/perylene was 800/1, SDS concentration was 150mmol/L and pH value above 9, according to the fluorescence quenching ,within a certain range of the concentration of Cu2+ion can be linearly determined.

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Self-assembly of Stimuli Responsive Coiled-coil Fibrous Hydrogels

Soft Matter ◽

10.1039/d1sm00780g ◽

2021 ◽

Author(s):

Michael Meleties ◽

Priya Katyal ◽

Bonnie Lin ◽

Dustin Britton ◽

Jin Kim Montclare

Keyword(s):

Drug Delivery ◽

Tissue Engineering ◽

Self Assembly ◽

Coiled Coil ◽

Stimuli Responsive ◽

Tunable Properties ◽

Stimuli Sensitive ◽

Biomedical Fields

Owing to their tunable properties, hydrogels comprised of stimuli sensitive polymers are one of the most appealing scaffolds with applications in tissue engineering, drug delivery and other biomedical fields. We...

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Identification of pilin pools in the membranes of Pseudomonas aeruginosa

Journal of Bacteriology ◽

10.1128/jb.152.2.687-691.1982 ◽

1982 ◽

Vol 152 (2) ◽

pp. 687-691

Author(s):

T H Watts ◽

E A Worobec ◽

W Paranchych

Keyword(s):

Staphylococcus Aureus ◽

Pseudomonas Aeruginosa ◽

Gel Electrophoresis ◽

Type Strain ◽

Wild Type Strain ◽

Polyacrylamide Gel Electrophoresis ◽

Protein A ◽

Sodium Dodecyl ◽

Wild Type ◽

Outer Membranes

The proteins of purified inner and outer membranes obtained from Pseudomonas aeruginosa strains PAK and PAK/2Pfs were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and treated with antiserum raised against pure pili. Bound antipilus antibodies were visualized by reaction with 125I-labeled protein A from Staphylococcus aureus. The results showed that there are pools of pilin in both the inner and outer membranes of P. aeruginosa and that the pool size in the multipiliated strain is comparable with that of the wild-type strain.

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Manuscript-C-dots assembly at interface-041020.pdf

10.26434/chemrxiv.12114063 ◽

2020 ◽

Author(s):

Amin Reza Zolghadr ◽

Behnam Rostami

Keyword(s):

Self Assembly ◽

Carbon Dots ◽

Sodium Dodecyl ◽

Surface Region ◽

Systematic Investigation ◽

Dynamics Simulation ◽

Surface Tension Gradient ◽

Bivariate Analysis ◽

Air Interface ◽

Sds Surfactant

We describe a systematic investigation of carbon dots (C-dots) assemblies fabricated at the liquid/air interface because of the surface tension gradient. This gradient is originally created by capillary action and increased by addition of sodium dodecyl sulfate (SDS) surfactant or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) phospholipid to the surface of C-dots aqueous mixture. The arrangement of carbon dots in liquid bulk phase (before self-assembly) and at the surface region (after self-assembly) was examined by TEM microscopy. The presence of SDS surfactant and POPC phospholipid at the air/water interface induced the C-dots compression. In addition, molecular dynamics simulation was conducted to obtain the structure of C-dots at liquid/vapor interface. The orientation of C-dots is evaluated quantitatively at water/vapor surface by using bivariate analysis.

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The gene encoding a major component of the lateral elements of synaptonemal complexes of the rat is related to X-linked lymphocyte-regulated genes

Molecular and Cellular Biology ◽

10.1128/mcb.14.2.1137-1146.1994 ◽

1994 ◽

Vol 14 (2) ◽

pp. 1137-1146

Author(s):

J H Lammers ◽

H H Offenberg ◽

M van Aalderen ◽

A C Vink ◽

A J Dietrich ◽

...

Keyword(s):

Amino Acid ◽

Sequence Similarity ◽

Sodium Dodecyl ◽

Coiled Coil ◽

Amino Acid Identity ◽

Male Rat ◽

Amino Acid Sequence Similarity ◽

Gene Encoding ◽

Synaptonemal Complexes ◽

Complex Protein

The lateral elements of synaptonemal complexes (SCs) of the rat contain major components with relative electrophoretic mobilities (M(r)S) of 30,000 and 33,000. After one-dimensional separation of SC proteins on polyacrylamide-sodium dodecyl sulfate gels, these components show up as two broad bands. These bands contain closely related proteins, as judged from their peptide maps and immunological reactivity. Using affinity-purified polyclonal anti-30,000- and anti-33,000-M(r) component antibodies, we isolated a cDNA encoding at least one of the 30,000- or 33,000-M(r) SC components. The protein predicted from the nucleotide sequence of the cDNA, called SCP3 (for synaptonemal complex protein 3), has a molecular mass of 29.7 kDa and a pI value of 9.4. It has a potential nucleotide binding site and contains stretches that are predicted to be capable of forming coiled-coil structures. In the male rat, the gene encoding SCP3 is transcribed exclusively in the testis. SCP3 has significant amino acid similarity to the pM1 protein, which is one of the predicted products of an X-linked lymphocyte-regulated gene family of the mouse: there are 63% amino acid sequence similarity and 35% amino acid identity between the SCP3 and pM1 proteins. However, SCP3 differs from pM1 in several respects, and whether the proteins fulfill related functions is still an open question.

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Preliminary Treatment of Urinary Proteins Improves Electrophoretic Analysis and Immunodetection

Clinical Chemistry ◽

10.1093/clinchem/38.6.860 ◽

1992 ◽

Vol 38 (6) ◽

pp. 860-863 ◽

Cited By ~ 8

Author(s):

J M Verdier ◽

B Dussol ◽

P Dupuy ◽

Y Berland ◽

J C Dagorn

Keyword(s):

Protein Pattern ◽

Protein A ◽

Sodium Dodecyl ◽

Complex Mixture ◽

Electrophoretic Analysis ◽

Renal Physiology ◽

Preliminary Treatment ◽

Urinary Proteins ◽

Simple Method ◽

Electrophoretic Migration

Abstract Analysis of urinary protein composition is an important tool in studies on renal physiology and physiopathology. Urine is, however, a complex mixture containing, besides protein, a variety of compounds such as salts, peptides, oligosaccharides, and glycosaminoglycans. Some of these compounds interfere with the electrophoretic migration of protein in sodium dodecyl sulfate-polyacrylamide gels and prevent correct analysis of the protein pattern. We describe a simple method for extracting urinary proteins that considerably improves their electrophoretic migration and subsequent immunodetection. This treatment involves ammonium sulfate fractionations (for precipitating proteins), EDTA (for inhibiting protein aggregation), and HCl hydrolysis (for removing glycosylaminoglycans). Recovery during extraction was found to be almost quantitative for total protein and three representative proteins: albumin, alpha 1-glycoprotein acid, and beta 2-microglobulin.

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