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191.
Rhodopsin is phosphorylated in a light-dependent manner by a kinase intrinsic to the rod outer segment. We have used chromatofocusing to separate six phosphorylated species of rhodopsin and have recovered in the pH gradient fractions 60-80% of the initial phosphorylated sample loaded on the column. The isolated species of rhodopsin coincide with the species that are observed in isoelectric focusing gels in the pH range 6.1-4.7. Unphosphorylated rhodopsin focuses at a pI of 6.0. Two species having two phosphates per rhodopsin with isoelectric points of 5.45 and 5.40 have been isolated. The phosphate to rhodopsin ratios for the remaining species are 3.8, 5.0, 6.1, and 8.2 with isoelectric points of 5.16, 4.99, 4.85, and 4.73, respectively. The chromatofocusing profile suggests that there may be multiple forms of rhodopsin with the same number of phosphates among some of the other phosphorylated forms of rhodopsin. 相似文献
192.
193.
B Gerber-Jenson A Kazin J M Kehoe C Scheffel B W Erickson G W Litman 《Journal of immunology (Baltimore, Md. : 1950)》1981,126(3):1212-1216
The amino acid sequence of the VH region of McE, a monoclonal IgM cryoimmunoglobulin, has been determined employing automated sequencing methodology. Digestion of the intact Fab mu component, derived by trypsin cleavage of the parent protein at elevated temperature with CNBr, followed by complete reduction and alkylation, yielded the intact light chain as well as the 2 CNBr fragments that constituted the VH. N-terminal sequencing of the larger unblocked CNBr fragment, along with a component fragment derived by cleavage by BNPS-Skatole, established the structure of the VH from position 88 through the V leads to C switch region. Citraconylation of the smaller, blocked fragment effected sufficient solubilization for enzymatic deblocking and direct sequencing of the N-terminal 20 residues of the VH. Complete trypsin digestion of the N-terminal CNBr fragment yielded 9 peptides that could be isolated by preparative cation exchange chromatography and gel filtration. The complete sequence of these peptides along with 4 chymotryptic peptides completed the primary structure of the VH region. The primary structure of McE appears to resemble that of He, previously identified as belonging to the VH II subgroup. The presence of characteristic CDR and FR regions as well as the identification of a probable site of glycosylation suggest that the cryoimmunoglobulin closely resembles noncryoglobulins in terms of overall structural composition. The cryoglobulin property may arise through alterations in individual residues or unfavorable arrangements of CDR and FR segments. 相似文献
194.
Felicia Gaskin David J. Litman Charles R. Cantor Michael L. Shelanski 《Journal of cellular biochemistry》1975,3(1):39-50
The porcine neurotubule and its basic subunit were found to be modified in vitro by iodination of amino acids (principally tyrosine) using lactoperoxidase. Iodide ion, H2O2, or lactoperoxidase singly or in any pairwise combination had virtually no effect on neurotubules. However, when all three reagents were present, permitting covalent iodination, it was found that at 0.1 iodotyrosines per tubulin dimer the microtubules unravel to form structures which morphologically resemble strands of protofilaments twisted or wound around each other. These abnormal tubules are stable at room temperature and 4°C. Both monomers of tubulin are labeled to approximately the same extent. Iodinated tubulin (0.1 iodotyrosines/dimer) is unable to assemble in vitro under normal assembly conditions. Heavily iodinated microtubules (8 iodines per tubulin dimer) are similar in morphology to the slightly iodinated structures. 相似文献
195.
196.
E Q Lawson B E Hedlund M E Ericson D A Mood G W Litman R Middaugh 《Archives of biochemistry and biophysics》1983,220(2):572-575
The effect of covalently attached carbohydrate on the solubility of a number of proteins has been examined by the PEG precipitation technique. Both increases and decreases in solubility are observed depending on the state of glycosylation, the type of protein, and temperature. It is concluded from this data and associated apparent thermodynamic parameters that a general role for carbohydrate in the solubilization of proteins is not currently experimentally supportable. 相似文献
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199.
Chewing of khat leaves has been noted to be widespread in Yemen. Immigrants to Israel brought that practice along and have kept it alive ever since their initial settlement over thirty years ago. The small epidemiological study reported here made an inquiry into the extent of khat use in two agricultural villages. It also explored the association of that practice with social and psychiatric variables. Of interest was the finding that — contrary to most addictions — the prevalence rate of psychopathology was not higher among users than among abstainers. 相似文献