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Reconstitution of the Vertebrate Visual Cascade Using Recombinant Heterotrimeric Transducin Purified from Sf9 Cells

Autor: K C Min, T P Sakmar, Stephen A. Gravina

Publikováno v: Protein Expression and Purification. 20:514-526

For reconstitution studies with rhodopsin and cGMP phosphodiesterase (PDE), all three subunits of heterotrimeric transducin (T alpha beta gamma) were simultaneously expressed in Sf9 cells at high levels using a baculovirus expression system and purif

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::93976d927c3a1754524d1167180beaf7
https://doi.org/10.1006/prep.2000.1326

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Transducin-alpha C-terminal mutations prevent activation by rhodopsin: a new assay using recombinant proteins expressed in cultured cells

Autor: Henry R. Bourne, Pablo Garcia, T. P. Sakmar, René Onrust, S. M. Bell

Publikováno v: The EMBO Journal. 14:4460-4469

We have measured the activation by recombinant rhodopsin of the alpha-subunit (alpha 1) of retinal transducin (Gt, also recombinant) using a new assay. Cultured cells are transiently transfected with DNAs encoding opsin and the three subunits of Gt (

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ba51c4ed605c814bea3398057254f28e
https://doi.org/10.1002/j.1460-2075.1995.tb00125.x

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Rapid activation of transducin by mutations distant from the nucleotide-binding site: evidence for a mechanistic model of receptor-catalyzed nucleotide exchange by G proteins

Autor: E P, Marin, A G, Krishna, T P, Sakmar

Publikováno v: The Journal of biological chemistry. 276(29)

G proteins act as molecular switches in which information flow depends on whether the bound nucleotide is GDP ("off") or GTP ("on"). We studied the basal and receptor-catalyzed nucleotide exchange rates of site-directed mutants of the alpha subunit o

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::26bdaace7b32950d8a7fe8e6f97e3069
https://pubmed.ncbi.nlm.nih.gov/11356823

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Assays for activation of recombinant expressed opsins by all-trans-retinals

Autor: M, Han, T P, Sakmar

Publikováno v: Methods in enzymology. 315

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::81264c7d4462e0213f40a71b86bdba73
https://pubmed.ncbi.nlm.nih.gov/10736707

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Structural determinants of active state conformation of rhodopsin: molecular biophysics approaches

Autor: K, Fahmy, T P, Sakmar, F, Siebert

Publikováno v: Methods in enzymology. 315

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::36bca7e66f2cf8fc2394032eba9dad35
https://pubmed.ncbi.nlm.nih.gov/10736702

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Analysis of functional microdomains of rhodopsin

Autor: S W, Lin, M, Han, T P, Sakmar

Publikováno v: Methods in enzymology. 315

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::4759aaa642e9055e82f4cb858356612f
https://pubmed.ncbi.nlm.nih.gov/10736698

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Evidence for the specific interaction of a lipid molecule with rhodopsin which is altered in the transition to the active state metarhodopsin II

Autor: M, Beck, F, Siebert, T P, Sakmar

Publikováno v: FEBS letters. 436(3)

Comparing the FTIR difference spectra of the rhodopsin --metarhodopsin II transition in membranes and in dodecylmaltoside detergent, characteristic variations are observed between 1715 and 1750 cm(-1). By repeating the measurements with the rhodopsin

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::230ebaef91d735eb7d44d2c99ab75ed2
https://pubmed.ncbi.nlm.nih.gov/9801137

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Mechanisms of spectral tuning in blue cone visual pigments. Visible and raman spectroscopy of blue-shifted rhodopsin mutants

Autor: S W, Lin, G G, Kochendoerfer, K S, Carroll, D, Wang, R A, Mathies, T P, Sakmar

Publikováno v: The Journal of biological chemistry. 273(38)

Spectral tuning by visual pigments involves the modulation of the physical properties of the chromophore (11-cis-retinal) by amino acid side chains that compose the chromophore-binding pocket. We identified 12 amino acid residues in the human blue co

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::4fe0490f107d888cf77c86d5cbec24ed
https://pubmed.ncbi.nlm.nih.gov/9733753

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Rhodopsin: a prototypical G protein-coupled receptor

Autor: T P, Sakmar

Publikováno v: Progress in nucleic acid research and molecular biology. 59

A variety of spectroscopic and biochemical studies of recombinant site-directed mutants of rhodopsin and related visual pigments have been reported over the past 9 years. These studies have elucidated key structural elements common to visual pigments

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::cc3e73bbbda637039884395f07c8a438
https://pubmed.ncbi.nlm.nih.gov/9427838

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Mutation of a conserved cysteine in the X-linked cone opsins causes color vision deficiencies by disrupting protein folding and stability

Autor: M A, Kazmi, T P, Sakmar, H, Ostrer

Publikováno v: Investigative ophthalmologyvisual science. 38(6)

To test the effects of disruption of a conserved cysteine in the green cone opsin molecule on light-activated isomerization, transducin activation, folding, transport, and protein half-life.Stable cell lines were established by transfecting 293-EBNA

Externí odkaz: https://explore.openaire.eu/search/publication?articleId=pmid________::ac26f7a2a4903e0e0578b6720b0b90bd
https://pubmed.ncbi.nlm.nih.gov/9152227

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