The transmembrane domain of glycophorin A as studied by cross-linking using photoactivatable phospholipids.

Ross, Alonzo H.; Radhakrishnan, R.; Robson, Robert J.; Khorana, H. G.

doi:10.1016/s0021-9258(18)34699-4

Cited by 102 publications

(35 citation statements)

References 53 publications

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“…However, carbon-hydrogen insertions appear to occur only in the absence of competing reactions . Electron-rich amino acids, such as tryptophan, glutamic acid, or cysteine are preferentially labeled relative to other amino acids Hoppe et al, 1983;Ross et al, 1982) . Furthermore, the lifetime of both the nitrene group, and the linear diazo isomer formed in high yields from the carbene precursor, allows for the modification of amino acids that are not in the immediate vicinity of the photoactivatable probe at the time of photolysis (Bayley and Knowles, 1978 ; .…”

Section: Inhibition Of Cross-linkmentioning

confidence: 99%

ER translocation intermediates are adjacent to a nonglycosylated 34-kD integral membrane protein.

Kellaris

Bowen

Gilmore

1991

The Journal of Cell Biology

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Abstract. We have used the homobifunctional crosslinking reagent disuccinimidyl suberate (DSS) to identify proteins that are adjacent to nascent polypeptides undergoing translocations across mammalian rough ER . Translocation intermediates were assembled by supplementing cell free translations of truncated mRNAs with the signal recognition particle (SRP) and microsomal membrane vesicles . Two prominent crosslinked products of 45 and 64 kD were detected . The 64-kD product was obtained when the cell free translation contained SRP, while formation of the 45-kD product required both SRP and translocation competent microsomal membrane vesicles . In agreement with previous investigators, we suggest that the 64-kD T HE signal sequence of proteins destined for translocatlon across or integration into the rough ER (RER)' contains the information that initiates sequential interactions with cytoplasmic and membrane bound components of the translocation apparatus . The cytoplasmic ribonucleoprotein signal recognition particle (SRP) cotranslationally selects proteins with RER signal sequences by binding to the nascent polypeptide when the signal sequence emerges from the large ribosomal subunit (Walter and Blobel, 19ß1b ;Walter and Blobel, 1982;Walter et al ., 1981) . The signal sequence recognition activity of SRP has been localized to the 54-kD subunit (SRP54) by photo-cross-linking experiments (Krieg et al., 1986;Kurzchalia et al., 1986) . The affinity of the SRP-ribosome complex for the membrane-bound SRP receptor (or docking protein) results in the selective delivery of the ribosome to the membrane surface (Gilmore et al., 19ß2b ; Meyer et al ., 19ß2a ; Walter and Blobel, 198 la), SRP receptor displaces SRP from the signal sequence in a GTPdependent reaction that allows the subsequent membrane insertion of the nascent polypeptide Gilmore and Blobel, 1983) . Although translocation of the polypeptide across the membrane bilayer is a poorly understood process, the preponderance of current experimental evidence suggests that nascent polypeptides undergoing transport are in contact with integral membrane proteins that 1. Abbreviations used in this paper: NEM, N-ethylmaleimide ; RER, rough endoplastnic reticulum; SRP, signal recognition particle .

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Section: Inhibition Of Cross-linkmentioning

confidence: 99%

ER translocation intermediates are adjacent to a nonglycosylated 34-kD integral membrane protein.

Kellaris

Bowen

Gilmore

1991

The Journal of Cell Biology

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“…The use of photoreactive membrane lipids to study lipid/peptide interactions is not entirely new. ,− Khorana et al performed pioneering work on the synthesis of phospholipids containing photoreactive precursors for studying lipid/protein interactions. − In this context, also azide-modified phospholipids were synthesized. ,, Although the azide moiety is less reactive than phenylazides and diazirines, ,, the azide modification has the advantage of less steric hindrance, enhanced stability, and facile incorporation into the fatty acid chain. Hence, it is conceivable that azide-modified lipids cause less disturbance of the lipid bilayer when they are mixed with bilayer-forming phospholipids.…”

Section: Introductionmentioning

confidence: 99%

Azide-Modified Membrane Lipids: Miscibility with Saturated Phosphatidylcholines

et al. 2019

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In this study, we describe the miscibility of four azide-modified membrane phospholipids (azidolipids) with conventional phospholipids. The azidolipids bear an azide group at different positions of the sn-1 or sn-2 alkyl chain and they further differ in the type of linkage (ester vs ether) of the sn-2 alkyl chain. Investigations regarding the miscibility of the azidolipids with bilayer-forming phosphatidylcholines will evaluate lipid mixtures that are suitable for the production of stable azidolipid-doped liposomes. These vesicles then serve as model membranes for the incorporation of model peptides or proteins in the future. The miscibility of both types of phospholipids was studied by calorimetric assays, electron microscopy, small-angle X-ray scattering, infrared spectroscopy, and dynamic light scattering to provide a complete biophysical characterization of the mixed systems.

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“…Further results of labeling using the phospholipid 31 showed that the Glu-70 residue is embedded in the membrane. 106 In addition, in combination with radiolabeling, two lipid analogues 34 and 35 containing TPD units ( Fig. 8 ) were designed to study the interactions between the influenza virus and target membrane lipids, 107 mitochondrial proteins and phosphatidylcholine.…”

Section: Applications Of Cmpal For Biological Molecule–protein Intera...mentioning

confidence: 99%

Current advances of carbene-mediated photoaffinity labeling in medicinal chemistry

Chen

et al. 2018

RSC Adv.

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Photoaffinity labeling (PAL) in combination with a chemical probe to covalently bind its target upon UV irradiation has demonstrated considerable promise in drug discovery for identifying new drug targets and binding sites. In particular, carbene-mediated photoaffinity labeling (cmPAL) has been widely used in drug target identification owing to its excellent photolabeling efficiency, minimal steric interference and longer excitation wavelength.Specifically, diazirines, which are among the precursors of carbenes and have higher carbene yields and greater chemical stability than diazo compounds, have proved to be valuable photolabile reagents in a diverse range of biological systems. This review highlights current advances of cmPAL in medicinal chemistry, with a focus on structures and applications for identifying small molecule-protein and macromolecule-protein interactions and ligand-gated ion channels, coupled with advances in the discovery of targets and inhibitors using carbene precursor-based biological probes developed in recent decades.

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The transmembrane domain of glycophorin A as studied by cross-linking using photoactivatable phospholipids.

Cited by 102 publications

References 53 publications

ER translocation intermediates are adjacent to a nonglycosylated 34-kD integral membrane protein.

ER translocation intermediates are adjacent to a nonglycosylated 34-kD integral membrane protein.

Azide-Modified Membrane Lipids: Miscibility with Saturated Phosphatidylcholines

Current advances of carbene-mediated photoaffinity labeling in medicinal chemistry

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