Measurement of the immune response to an allograft: Amino acid incorporation by peripheral white blood cells in man

Abstract The study of maturation of rabbit reticulocytes in vivo revealed an essential correlation between the degree of reticulocytosis, the RNA content of the circulating red blood cells and their ability to carry out amino acid incorporation into protein. The nevertheless frequent occurrence of considerable quantitative discrepancies between these parameters during the progress of maturation appears to be attributable to fluctuations in the reticulocyte population due to intrinsic difference in the life span of these cells and to release of young reticulocytes from the hyperplastic bone marrow into the circulation. The distortions in the pattern of maturation kinetics by the continued contribution of the bone marrow seem to be largely eliminated by blocking erythropoiesis at the height of reticulocytosis with the aid of actinomycin D treatment.

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A STABILIZED ENZYME SYSTEM FOR AMINO ACID INCORPORATION

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)70686-8 ◽

1957 ◽

Vol 228 (1) ◽

pp. 23-39

Author(s):

Howard Sachs

Keyword(s):

Amino Acid ◽

Enzyme System ◽

Amino Acid Incorporation ◽

Acid Incorporation

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AMINO ACID INCORPORATION BY INTACT AND DISRUPTED RIBONUCLEOPROTEIN PARTICLES

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)70638-8 ◽

1957 ◽

Vol 229 (1) ◽

pp. 535-546

Author(s):

George C. Webster

Keyword(s):

Amino Acid ◽

Amino Acid Incorporation ◽

Acid Incorporation ◽

Ribonucleoprotein Particles

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Transferability of N-terminal mutations of pyrrolysyl-tRNA synthetase in one species to that in another species on unnatural amino acid incorporation efficiency

Amino Acids ◽

10.1007/s00726-020-02927-z ◽

2020 ◽

Author(s):

Thomas L. Williams ◽

Debra J. Iskandar ◽

Alexander R. Nödling ◽

Yurong Tan ◽

Louis Y. P. Luk ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Genetic Code ◽

Unnatural Amino Acids ◽

Trna Synthetase ◽

Unnatural Amino Acid ◽

Amino Acid Incorporation ◽

Acid Incorporation ◽

Genetic Code Expansion ◽

Incorporation Efficiency

AbstractGenetic code expansion is a powerful technique for site-specific incorporation of an unnatural amino acid into a protein of interest. This technique relies on an orthogonal aminoacyl-tRNA synthetase/tRNA pair and has enabled incorporation of over 100 different unnatural amino acids into ribosomally synthesized proteins in cells. Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA from Methanosarcina species are arguably the most widely used orthogonal pair. Here, we investigated whether beneficial effect in unnatural amino acid incorporation caused by N-terminal mutations in PylRS of one species is transferable to PylRS of another species. It was shown that conserved mutations on the N-terminal domain of MmPylRS improved the unnatural amino acid incorporation efficiency up to five folds. As MbPylRS shares high sequence identity to MmPylRS, and the two homologs are often used interchangeably, we examined incorporation of five unnatural amino acids by four MbPylRS variants at two temperatures. Our results indicate that the beneficial N-terminal mutations in MmPylRS did not improve unnatural amino acid incorporation efficiency by MbPylRS. Knowledge from this work contributes to our understanding of PylRS homologs which are needed to improve the technique of genetic code expansion in the future.

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