Identification and Semiquantitative Determination of Phenyl Thiohydantoins.

Edman, Pehr; Sjöquist, John; Lunde, K.; Eliasson, N. A.; Thorell, B.

doi:10.3891/acta.chem.scand.10-1507

Cited by 307 publications

(53 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further, the NH2-terminal region can be expected to be external to the viral envelope by comparison with other viral (21) and nonviral (17,22) E2 is known to be external to the viral envelope, no evidence of carbohydrate attachment is seen at least through the first 40 residues of each protein. This is in contrast to the situation found in glycophorin (15) in which carbohydrate is attached to the second residue and there are 16 carbohydrate attachment sites in the first 50 residues. In many virus systems, the production of functional structural proteins involves proteolytic cleavage (23).…”

Section: Resultscontrasting

confidence: 86%

See 1 more Smart Citation

Amino-terminal sequence analysis of the structural proteins of Sindbis virus.

Bell¹,

Hunkapiller²,

Hood³

et al. 1978

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The structural proteins of Sindbis virus, an enveloped virus which belongs to the Togavirus family, have been subjected to automated Edman degradation using improved techniques. Extensive NHrterminal sequences of about 50 residues were determined for each of the two membrane glycoproteins. In both cases the NH2 terminus of the molecule was found to be similar in composition to typical water-soluble proteins. The viral capsid protein was found to have a blocked a-amino group. This is consistent with other observations that viral proteins derived from the NH2 terminus of precursor molecules are often blocked.Sindbis virus is a simple enveloped virus which matures by budding through a modified host cell plasma membrane (1). The virus particle contains only three structural proteins (2). One of these, the capsid protein, can be isolated from virus particles in the form of a nucleocapsid structure also containing the viral RNA (3). In the intact virus particle, this nucleocapsid is surrounded by a lipid envelope, largely in the form of a bilayer (4). Associated with the envelope are the other two structural proteins, glycoproteins El and E2. Although most of their mass is external to the lipid bilayer, they have the properties of integral membrane proteins. Mild detergent treatment is required to separate El and E2 from the other components of the virus (3,5), and at least one is a transmembrane protein because it extends completely through the bilayer and can be crosslinked to the capsid protein by suitable reagents (6).It has been shown that all three Sindbis structural proteins as well as those of the closely related Semliki Forest virus are formed by cleavage of a single precursor polypeptide (7-9), probably while the polypeptide chain is still nascent. The capsid protein is released into the cytoplasm of the cell; in contrast, El and E2 are inserted into the lumen of the rough endoplasmic reticulum (10). At some time after synthesis, they appear glycosylated at the cell plasmalemma, where they diffuse freely over the surface of the cell (11). The budding process appears to involve an interaction between preformed nucleocapsids and that region of one or both of the envelope proteins exposed at the cytoplasmic face of the plasmalemma. This MATERIALS AND METHODS Protein Preparation. The growth and purification of Sindbis virus and the separation of the structural proteins will be described in detail elsewhere. Briefly, the HR strain of Sindbis virus, grown at low multiplicity of infection in primary chicken embryo fibroblasts, was concentrated from culture fluid by precipitation with polyethylene glycol followed by pelleting onto a fluorocarbon cushion. The resuspended virus was then purified by sucrose gradient velocity centrifugation followed by isopycnic centrifugation in sucrose/2H20 gradients. These steps of the procedure were similar to those described previously (14). After disruption with Triton X-100, capsids were removed by centrifugation and the two envelope glycoproteins were separated by pas...

show abstract

Section: Resultscontrasting

confidence: 86%

“…1, and (Fig. 3), the lag increased (and repetitive yields decreased slightly) near cycle 15 The relative occurrence of each group of amino acids is expressed on a mol % basis. Data for the NH2-terminal regions of the proteins were computed from Fig.…”

Section: Resultsmentioning

confidence: 97%

Amino-terminal sequence analysis of the structural proteins of Sindbis virus.

Bell¹,

Hunkapiller²,

Hood³

et al. 1978

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…For certain residues, the phenylthiohydantoin derivatives were acid-hydrolyzed (30), and the free amino acids were identified on a Beckman model 121 amino-acid analyzer. Other phenylthiohydantoin derivatives were identified by a modification of the thin-layer chromatography system of Edman and Sjoquist (31). Generally, 6% of the phenylthiohydantoin derivative was subjected to gas chromatography, 50% to thin layer chromatography, and, when necessary, the remaining 44% was hydrolyzed and subjected to amino-acid analysis on an amino-acid analyzer.…”

Section: Methodsmentioning

confidence: 99%

Light Chain Sequences of Human IgM Cold Agglutinins

et al. 1972

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The amino-terminal amino-acid sequence has been determined for the kappa light chains of nine IgM cold agglutinins with specificity for blood grouprelated carbohydrate antigens. Six of the nine sequences corresponded to the prototype VKIII subgroup pattern, two to that of subgroup VKII, and only one to the VKI subgroup. This distribution of kappa subgroups differs markedly from that of normal and myeloma proteins sequenced to date, where the VKI subgroup is more prevalent than either VKii or VKiii. Evidence is presented that supports the conclusion that the unusual subgroup distribution relates to the antibody property itself and not to some other attribute of these molecules.

show abstract

“…The second method was the Edman degradation performed by the paper strip method of FraenkelConrat et al [26], followed by thin-layer chromatography on silica gel using the solvent system of Edman and Sjoquist [27,28].…”

Section: Amino-terminal End-group Analysismentioning

confidence: 99%

Octopine Dehydrogenase

Olomucki

et al. 1972

European Journal of Biochemistry

View full text Add to dashboard Cite

The amino acid composition of octopine dehydrogenase from muscles of Pecten maximus was determined. The calculate dpartial specifk volume of 0.74 allowed us to evaluate the molecular weight of the enzyme as 38000.It is concluded that this enzyme consists of a single polypeptide chain on the basis of the molecular weight determined from equilibrium ultracentrifugation in 6 M guanidine hydrochloride and from electrophoresis on polyacrylamide gel containing sodium dodecylsulfate as well as on the basis of the number of peptides obtained by tryptic digestion. The presence of one essential -SHgroup per mole of enzyme and of one binding site for NADH are also consistent with the existence of a single active center.No NH,-terminal amino acid can be detected by reaction with dansyl chloride and with phenylisothiocyanate.Controlled carboxypeptidase action showed that the C-terminal amino acid is lysine.

show abstract

Identification and Semiquantitative Determination of Phenyl Thiohydantoins.

Cited by 307 publications

References 0 publications

Amino-terminal sequence analysis of the structural proteins of Sindbis virus.

Amino-terminal sequence analysis of the structural proteins of Sindbis virus.

Light Chain Sequences of Human IgM Cold Agglutinins

Octopine Dehydrogenase

Contact Info

Product

Resources

About