Phosphoproteins in dictyostelium discoideum

Coffman, D.S.; Leichtling, Ben H.; Rickenberg, H.V.

doi:10.1002/jsscb.1981.380150407

Cited by 40 publications

(14 citation statements)

References 37 publications

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“…At the same time, the preceding discussion has assumed that the observed changes were due to altered polypeptide synthesis, and this need not be the case for all species. Coffman et al (33) have reported developmental changes in phosphorylation of plasma membrane proteins, and we (40) have previously reported that the processing at N-linked oligosaccharides undergoes a sharp transition at the tip formation stages. In addition to these posttranslational modifications, another possibility is the binding of pre-existing proteins to newly synthesized membrane receptor sites.…”

Section: )mentioning

confidence: 60%

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Das¹,

Henderson²

1983

The Journal of Cell Biology

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Developmental changes in the plasma membrane proteins of Dictyostelium discoideum have been studied using metabolic labeling with []SS]methionine and two-dimensional electrophoresis. Pulse labeling for 1 h at the early interphase, late interphase, aggregation, and tip formation stages of development showed that the profile of newly synthesized plasma membrane proteins changed dramatically over this interval. Only 14% of the polypeptide species were synthesized at all four stages at detectable levels; 86% of the species changed over this developmental interval according to the criterion that they were synthesized at some but not all of the four stages tested. Long-term labeling during vegetative growth followed by initiation of development showed that the "steady-state" levels of the plasma membrane proteins changed very little over the same period. The only changes were in minor species (33% overall change). Similar analyses of whole cell proteins showed 27 and 20% change, respectively. Cell surface radioiodination revealed 52 external proteins in the plasma membrane. Comparison with the uniform methionine labeling results showed that these proteins were, with one notable exception, minor membrane components. In these external proteins, also, developmental changes were limited and were observed in the less abundant species.These results demonstrate the existence of two general classes of plasma membrane proteins. The first is a population of high-abundance proteins that are present in vegetative cells and are largely conserved through development. These possibly serve "housekeeping" functions common to all stages. The second class consists of low-abundance species that are expressed in a highly stage-specific manffer and which presumably participate in developmentally important functions.

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Section: )mentioning

confidence: 60%

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Das¹,

Henderson²

1983

The Journal of Cell Biology

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“…gp8O is known to be highly glycosylated (22) and phosphorylated (4,29). Digestion of the purified protein with periodate abolished its ability to react (after transfer to nitrocellulose) with either monoclonal antibody E28D8 or the polyclonal rabbit antibodies (Fig.…”

Section: Resultsmentioning

confidence: 98%

“…A series of investigations by Gerisch and his colleagues (1)(2)(3)(4) has demonstrated that the formation of these adhesions, which define "contact sites A," can be prevented by Fab antibody fragments reactive with cell surface components. A cell surface glycoprotein with an apparent molecular weight of 80,000 (gp8O) has been identified as a major species which reacts with these Fab fragments (21,22).…”

mentioning

confidence: 99%

Monoclonal antibody recognizing gp80, a membrane glycoprotein implicated in intercellular adhesion of Dictyostelium discoideum.

Murray¹,

Niman²,

Loomis³

1983

Mol. Cell. Biol.

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We have raised a monoclonal antibody, designated E28D8, which reacts with an 80,000-dalton membrane glycoprotein (gp8O) of Dictyostelium discoideum. gp8O has been implicated in the formation of the EDTA-resistant adhesions ("contact sites A") which appear during development. The monoclonal antibody reacted with other developmentally regulated proteins of D. discoideum, confirming previous results indicating the presence of common antigenic determinants recognized by polyclonal rabbit antibodies directed to gp8O. Periodate sensitivity of the determinants suggests that carbohydrate may be necessary for reactivity. Thus, the determinant recognized by E28D8 may result from a posttranslational modification common to a number of proteins. Some of the proteins that carry the determinant were preferentially localized to posterior cells in slugs. Monoclonal antibody E28D8 did not inhibit contact-sites-A-mediated intercellular adhesion. However, gp8O affinity purified on immobilized monoclonal antibody was able to neutralize the adhesion-blocking effect of rabbit antiserum to gp8O. Although gp8O itself may not be essential for cell-cell adhesion, it appears to carry the determinants associated with adhesion.

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“…With 50 pM [Y-~,P]ATP, the apparent Km's for MBP and histone H1 were 70 and 700pg/ml, respectively. For MBP and histone H1 phosphorylation, optimal activity was observed with 7 mM MgC12 in the presence of 5 mM EGTA and 2 mM EDTA.…”

Section: Characterization Of the 300-400 Kda Protein Kinase Activitymentioning

confidence: 99%

“…In addition, a CaZf/calmodulin-insensitive myosin light chain kinase has been partially purified and characterized (1 1). These protein kinases are all relatively specific for myosin, and are unlikely to account for all of the many changes in protein phosphorylation that accompany development (7).…”

Section: Introductionmentioning

confidence: 99%

Characterization of Developmentally Regulated cAMP/Ca²⁺‐Independent Protein Kinases from Dictyostelium discoideum

1989

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Cell-free extracts of the slime mold Dictyostelium discoideum were assayed for phosphorylating activity towards endogenous proteins and towards histone H1, casein and myelin basic protein (MBP). During development, protein kinase activity towards all of these substrates steadily increased and peaked between the aggregation and the pseudoplasmodial stages. Particulate-associated kinase activity was solubilized with 1% CHAPS, and separated into 300-400 kDa and -100 kDa components on Sephacryl S-300. The 300-400 kDa peak exhibited the most pronounced developmental increase in MBP phosphorylating activity. It was further fractionated on DEAE-Sephacel and heparin-Sepharose, and in each case, it coeluted with the histone H1 phosphorylating activity. The activity of this kinase was unaffected by cAMP and calmodulin, but it was reduced to 50% by -350 mM NaCI, 5 mM NaF and 4 0~ polylysine/ml. The -100 kDa peak exhibited the most pronounced increase in casein kinase activity during development. Most of the casein phosphorylating activity did not bind to DEAE-Sephacel; it was distinct from casein kinase 2, which was not developmentally regulated. In parallel with these elevated kinase activities during development, there was increased in vitro phosphorylation of a number of Dictyostelium proteins, including two major phosphoproteins of 140 and 94 kDa.

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Phosphoproteins in dictyostelium discoideum

Cited by 40 publications

References 37 publications

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Monoclonal antibody recognizing gp80, a membrane glycoprotein implicated in intercellular adhesion of Dictyostelium discoideum.

Characterization of Developmentally Regulated cAMP/Ca²⁺‐Independent Protein Kinases from Dictyostelium discoideum

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Phosphoproteins in dictyostelium discoideum

Cited by 40 publications

References 37 publications

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Developmental regulation of Dictyostelium discoideum plasma membrane proteins.

Monoclonal antibody recognizing gp80, a membrane glycoprotein implicated in intercellular adhesion of Dictyostelium discoideum.

Characterization of Developmentally Regulated cAMP/Ca2+‐Independent Protein Kinases from Dictyostelium discoideum

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Characterization of Developmentally Regulated cAMP/Ca²⁺‐Independent Protein Kinases from Dictyostelium discoideum