Retention of the cis proline conformation in tripeptide fragments of bovine pancreatic ribonuclease A containing a non-natural proline analogue, 5,5-dimethylproline

Seong Soo A An, Cathy C. Lester, Jin Lin Peng, Yue Jin Li, David M. Rothwarf, E. Welker, Theodore W. Thannhauser, L. S. Zhang, James P. Tam, Harold A. Scheraga

Research output: Contribution to journalArticle

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Abstract

Attention is focused on L-5,5-dimethylproline (dmP) as a substitute to lock L-proline (Pro) in a cis conformation in peptides and proteins, to prevent cis/trans isomerization when a protein with cis X-Pro peptide groups unfolds. Procedures have been developed to obtain optically pure L-dmP and to incorporate this sterically hindered residue as the central one in tripeptides that are suitable for fragment coupling to prepare synthetic proteins. Based on the sequences of residues 92-94 (Tyr-Pro-Asn:YPN) and 113- 115 (Asn-Pro-Tyr:NPY) in bovine pancreatic ribonuclease A (RNase A), in which the X-Pro peptide groups are in the cis conformation, the tripeptides Ac-Tyr- dmP-Asn (YdmPN) and Ac-Asn-dmP-Tyr (NdmPY) were synthesized, and their structures were determined by 2D 1H nuclear magnetic resonance (NMR) spectroscopy. YdmPN was found to exist solely in the cis conformation between 6 and 60 °C, whereas NdmPY was found to have some trans component that increased from about 10% to about 21% as the temperature increased over the range between 6 and 80 °C. Both YdmPN and cis-NdmPY adopt a type VI reverse turn, as does proline. The NMR structures of YdmPN and cis-NdmPY are comparable with the X-ray structures of the corresponding portions of RNase A, and the NMR structure of trans-NdmPY is compatible with the X-ray structure of the isolated tripeptide, Ac-NPY. These results demonstrate that L-dmP is a promising substitute for proline in a variety of protein problems to constrain the X-Pro peptide group to the cis conformation.

Original languageEnglish
Pages (from-to)11558-11566
Number of pages9
JournalJournal of the American Chemical Society
Volume121
Issue number49
DOIs
Publication statusPublished - Dec 15 1999

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Pancreatic Ribonuclease
Proline
Peptides
Conformations
Proteins
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
X-Rays
X rays
Isomerization
Nuclear magnetic resonance spectroscopy
Temperature
stachydrine
Ribonucleases

ASJC Scopus subject areas

  • Chemistry(all)

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Retention of the cis proline conformation in tripeptide fragments of bovine pancreatic ribonuclease A containing a non-natural proline analogue, 5,5-dimethylproline. / An, Seong Soo A; Lester, Cathy C.; Peng, Jin Lin; Li, Yue Jin; Rothwarf, David M.; Welker, E.; Thannhauser, Theodore W.; Zhang, L. S.; Tam, James P.; Scheraga, Harold A.

In: Journal of the American Chemical Society, Vol. 121, No. 49, 15.12.1999, p. 11558-11566.

Research output: Contribution to journalArticle

An, Seong Soo A ; Lester, Cathy C. ; Peng, Jin Lin ; Li, Yue Jin ; Rothwarf, David M. ; Welker, E. ; Thannhauser, Theodore W. ; Zhang, L. S. ; Tam, James P. ; Scheraga, Harold A. / Retention of the cis proline conformation in tripeptide fragments of bovine pancreatic ribonuclease A containing a non-natural proline analogue, 5,5-dimethylproline. In: Journal of the American Chemical Society. 1999 ; Vol. 121, No. 49. pp. 11558-11566.
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abstract = "Attention is focused on L-5,5-dimethylproline (dmP) as a substitute to lock L-proline (Pro) in a cis conformation in peptides and proteins, to prevent cis/trans isomerization when a protein with cis X-Pro peptide groups unfolds. Procedures have been developed to obtain optically pure L-dmP and to incorporate this sterically hindered residue as the central one in tripeptides that are suitable for fragment coupling to prepare synthetic proteins. Based on the sequences of residues 92-94 (Tyr-Pro-Asn:YPN) and 113- 115 (Asn-Pro-Tyr:NPY) in bovine pancreatic ribonuclease A (RNase A), in which the X-Pro peptide groups are in the cis conformation, the tripeptides Ac-Tyr- dmP-Asn (YdmPN) and Ac-Asn-dmP-Tyr (NdmPY) were synthesized, and their structures were determined by 2D 1H nuclear magnetic resonance (NMR) spectroscopy. YdmPN was found to exist solely in the cis conformation between 6 and 60 °C, whereas NdmPY was found to have some trans component that increased from about 10{\%} to about 21{\%} as the temperature increased over the range between 6 and 80 °C. Both YdmPN and cis-NdmPY adopt a type VI reverse turn, as does proline. The NMR structures of YdmPN and cis-NdmPY are comparable with the X-ray structures of the corresponding portions of RNase A, and the NMR structure of trans-NdmPY is compatible with the X-ray structure of the isolated tripeptide, Ac-NPY. These results demonstrate that L-dmP is a promising substitute for proline in a variety of protein problems to constrain the X-Pro peptide group to the cis conformation.",
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T1 - Retention of the cis proline conformation in tripeptide fragments of bovine pancreatic ribonuclease A containing a non-natural proline analogue, 5,5-dimethylproline

AU - An, Seong Soo A

AU - Lester, Cathy C.

AU - Peng, Jin Lin

AU - Li, Yue Jin

AU - Rothwarf, David M.

AU - Welker, E.

AU - Thannhauser, Theodore W.

AU - Zhang, L. S.

AU - Tam, James P.

AU - Scheraga, Harold A.

PY - 1999/12/15

Y1 - 1999/12/15

N2 - Attention is focused on L-5,5-dimethylproline (dmP) as a substitute to lock L-proline (Pro) in a cis conformation in peptides and proteins, to prevent cis/trans isomerization when a protein with cis X-Pro peptide groups unfolds. Procedures have been developed to obtain optically pure L-dmP and to incorporate this sterically hindered residue as the central one in tripeptides that are suitable for fragment coupling to prepare synthetic proteins. Based on the sequences of residues 92-94 (Tyr-Pro-Asn:YPN) and 113- 115 (Asn-Pro-Tyr:NPY) in bovine pancreatic ribonuclease A (RNase A), in which the X-Pro peptide groups are in the cis conformation, the tripeptides Ac-Tyr- dmP-Asn (YdmPN) and Ac-Asn-dmP-Tyr (NdmPY) were synthesized, and their structures were determined by 2D 1H nuclear magnetic resonance (NMR) spectroscopy. YdmPN was found to exist solely in the cis conformation between 6 and 60 °C, whereas NdmPY was found to have some trans component that increased from about 10% to about 21% as the temperature increased over the range between 6 and 80 °C. Both YdmPN and cis-NdmPY adopt a type VI reverse turn, as does proline. The NMR structures of YdmPN and cis-NdmPY are comparable with the X-ray structures of the corresponding portions of RNase A, and the NMR structure of trans-NdmPY is compatible with the X-ray structure of the isolated tripeptide, Ac-NPY. These results demonstrate that L-dmP is a promising substitute for proline in a variety of protein problems to constrain the X-Pro peptide group to the cis conformation.

AB - Attention is focused on L-5,5-dimethylproline (dmP) as a substitute to lock L-proline (Pro) in a cis conformation in peptides and proteins, to prevent cis/trans isomerization when a protein with cis X-Pro peptide groups unfolds. Procedures have been developed to obtain optically pure L-dmP and to incorporate this sterically hindered residue as the central one in tripeptides that are suitable for fragment coupling to prepare synthetic proteins. Based on the sequences of residues 92-94 (Tyr-Pro-Asn:YPN) and 113- 115 (Asn-Pro-Tyr:NPY) in bovine pancreatic ribonuclease A (RNase A), in which the X-Pro peptide groups are in the cis conformation, the tripeptides Ac-Tyr- dmP-Asn (YdmPN) and Ac-Asn-dmP-Tyr (NdmPY) were synthesized, and their structures were determined by 2D 1H nuclear magnetic resonance (NMR) spectroscopy. YdmPN was found to exist solely in the cis conformation between 6 and 60 °C, whereas NdmPY was found to have some trans component that increased from about 10% to about 21% as the temperature increased over the range between 6 and 80 °C. Both YdmPN and cis-NdmPY adopt a type VI reverse turn, as does proline. The NMR structures of YdmPN and cis-NdmPY are comparable with the X-ray structures of the corresponding portions of RNase A, and the NMR structure of trans-NdmPY is compatible with the X-ray structure of the isolated tripeptide, Ac-NPY. These results demonstrate that L-dmP is a promising substitute for proline in a variety of protein problems to constrain the X-Pro peptide group to the cis conformation.

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