Ultrasensitive silver-stain method for the detection of proteins in polyacrylamide gels and immunoprecipitates on agarose gels

Porro, Massimo; Viti, S; Antoni, G.

doi:10.1016/0003-2697(82)90179-8

Cited by 48 publications

(13 citation statements)

References 10 publications

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“…Entire samples were electrophoresed on 7.5% SDS-polyacrylamide gels (Laemmli, 1970) until a 116-kD prestained marker (P-galactosidase) was run 4.5 to 5 cm into the separating gel. Gels were then silvei stained (Porro et al, 1982), dried, and autoradiographed.…”

Section: Microsomal Membrane Preparation In Vitro Phosphorylationmentioning

confidence: 99%

Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli

Liscum¹,

Briggs²

1995

The Plant Cell

136

236

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Section: Microsomal Membrane Preparation In Vitro Phosphorylationmentioning

confidence: 99%

Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli

Liscum¹,

Briggs²

1995

The Plant Cell

136

236

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“…Samples were diluted with onehalf volume of sample running buffer (5% SDS, 25% sucrose, 0.05% bromophenol blue) and boiled for 1 min before separation by SDSPAGE, as described by Laeininli (1970), on 7.5-15% polyacrylamide gradient gels. Polypeptides were visualized by silver staining (Porro et al 1982). Approximately 5 pg of each protein sample was loaded per lane for all onedimensional gels.…”

Section: Specimens Of Amentioning

confidence: 99%

DIFFERENCES IN THE PROTEIN PROFILES OF CULTURED AND ENDOSYMBIOTIC SYMBIODINIUM SP. (PYRROPHYTA) FROM THE ANEMONE AIPTASIA PALLIDA (ANTHOZOA)

Stochaj

Grossman

1997

Journal of Phycology

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One-and twedimensional sodium dodecylsulfate-polyacly lamide gel electrophoresis and immunological analyses were used to visualize dijferences i n polypeptides synthesized by Symbiodinium sp. from the anemone Aiptasia pallida when grown i n the cultured and endosymbiotic states (freshly isolated zooxanthellae). Surprisingly, a comparison of proteins in cultured and endosymbiotic Symbiodinium sp. revealed only four major polypeptides with similar isoelectric and molecular mass characteristics. Using monospeci$c antibodies, we demonstrated differences in .spec@ proteins synthesized lq the dinoflagellate in the two dgerent growth states. The dimeric, 14 kDa f m of the peripheral membrane peridinin-chlorophyll a binding pre tein predominates under endosymbiotic conditions, whereas the mon.omm'c, 35 kDa fm predominates under the culture conditions used i n this study. Antibodies to form II ribulose-1,5-bisphosphate carboxylase revealed 62 and 60 kDa ,forms of this protein i n the alga grown as a n e n d e symbiont and in, culture, respectively. Differences in the integral membrane p~~i n i n -c h l~o p h y l l a-c-binding proteins were also observed. These results demonstrate that there are major changes in the populations of proteins synthesized by Symbiodinium sp. in response to the conditions in hospite. Such changes may reflect a developmental switch that tailors the physiology of the alga to the conditions encountered in the endosymbiotic state.Kqy index words: dqferential protein expression; peridinin-chlorophyll a-binding proteins; peridinin-chlorophyll a/c-binding proteins; ribulose-1,5-bisphosphate carboxyluse; symbiosis; two-dimensional gel electrophoresis; western blot analysisIn the marine environment, symbiotic associations occur between photosynthetic algae and more than 250 species of nonphotosynthetic organisms, including cnidarians, sponges, platyhelminths, ascidians, molluscs, and protozoans (reviewed in Trench 1993). Of these, perhaps the best studied is the association between cnidarians and their dinoflagellate endosymbionts, the majority of which are of the genus Symbiodinium (Trench 1993). These dinoflagellates are members of the group of endosymbiotic algae commonly called zooxanthellae. In algal/cnidarian symbioses, zooxanthellae contribute to the growth and maintenance of their hosts by providing photosynthetically fixed carbon (reviewed in Trench

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“…Polypeptides were visualized by silver staining (Porro et al, 1982). To detect glycosylated proteins, periodic acid-Schiff staining was performed as described by Thornton et al (1994).…”

Section: Sds-pagementioning

confidence: 99%

Biochemical Characterization of the Extracellular Phosphatases Produced by Phosphorus-Deprived Chlamydomonas reinhardtii

1996

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We have examined the extracellular phosphatases produced by the terrestrial green alga Chlamydomonas reinbardfii in response to phosphorus deprivation. Phosphorus-deprived cells increase extracellular alkaline phosphatase activity 300-fold relative to unstarved cells. The alkaline phosphatases are released into the medium by cell-wall-deficient strains and by wild-type cells after treatment with autolysin, indicating that they are localized to the periplasm. Anion-exchange chromatography and analysis by nondenaturing polyacrylamide gel electrophoresis revealed that there are two major inducible alkaline phosphatases. A calcium-dependent enzyme composed of 190-kD glycoprotein subunits accounts for 85 to 95% of the alkaline phosphatase activity. lhis phosphatase has optimal activity at pH 9.5 and a K,,, of 120 to 262 PM for all physiological substrates tested, with the exception of phytic acid, which it cleaved with a 50-fold lower efficiency. An enzyme with optimal activity at pH 9 and no requirement for divalent cations accounts for 2 to 10% of the alkaline phosphatase activity. This phosphatase was only able to efficiently hydrolyze arylphosphates. l h e information reported here, in conjunction with the results of previous studies, defines the complement of extracellular phosphatases produced by phosphorus-deprived Chlamydomonas cells.Phosphorus is often a limiting nutrient in both terrestrial and aquatic ecosystems. Most organisms acquire phosphorus through active transport of Pi from the surrounding medium. However, in many soils more than 50% of the soluble phosphate is esterified to organic compounds, rendering it impossible for most organisms to absorb directly (Halstead and McKercher, 1975). Esterified Pi can be released from organic phosphates by the action of extracellular phosphatases, which catalyze the hydrolysis of the

show abstract

Ultrasensitive silver-stain method for the detection of proteins in polyacrylamide gels and immunoprecipitates on agarose gels

Cited by 48 publications

References 10 publications

Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli

Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli

DIFFERENCES IN THE PROTEIN PROFILES OF CULTURED AND ENDOSYMBIOTIC SYMBIODINIUM SP. (PYRROPHYTA) FROM THE ANEMONE AIPTASIA PALLIDA (ANTHOZOA)

Biochemical Characterization of the Extracellular Phosphatases Produced by Phosphorus-Deprived Chlamydomonas reinhardtii

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