Detection of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins by sensitivity- enhanced NMR spectroscopy

Aizhou Liu; Weidong Hu; Ananya Majumdar; Michael K. Rosen; Dinshaw J. Patel

doi:10.1023/A:1008373501591

Detection of very weak side chain-main chain hydrogen bonding interactions in medium-size ¹³C/¹⁵N-labeled proteins by sensitivity- enhanced NMR spectroscopy

Aizhou Liu, Weidong Hu, Ananya Majumdar, Michael K. Rosen, Dinshaw J. Patel

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

We describe the direct observation of very weak side chain-main chain hydrogen bonding interactions in medium-size ¹³C/¹⁵N-labeled proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the hydrogen acceptor side chain carboxylate carbon ¹³CO₂(δ) of glutamate 54 and the hydrogen donor backbone amide ¹⁵N of methionine 49 in a 12 kDa protein, human FKBP12, is detected via the trans- hydrogen bond (3h)J(NCO2δ) coupling by employing a novel sensitivity- enhanced HNCO-type experiment, CPD-HNCO. The (3h)J(NCO2δ) coupling constant appears to be even smaller than the average value of backbone (3h)J(NC') couplings, consistent with more extensive local dynamics in protein side chains.

Original language	English (US)
Pages (from-to)	79-82
Number of pages	4
Journal	Journal of biomolecular NMR
Volume	17
Issue number	1
DOIs	https://doi.org/10.1023/A:1008373501591
State	Published - 2000

Keywords

FKBP12
NMR detection
Sensitivity enhancement
Side chain-main chain hydrogen bonds

ASJC Scopus subject areas

Biochemistry
Spectroscopy

Access to Document

10.1023/A:1008373501591

Cite this

@article{4c710282bb9b479195496a16e5a8cc39,

title = "Detection of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins by sensitivity- enhanced NMR spectroscopy",

abstract = "We describe the direct observation of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the hydrogen acceptor side chain carboxylate carbon 13CO2(δ) of glutamate 54 and the hydrogen donor backbone amide 15N of methionine 49 in a 12 kDa protein, human FKBP12, is detected via the trans- hydrogen bond (3h)J(NCO2δ) coupling by employing a novel sensitivity- enhanced HNCO-type experiment, CPD-HNCO. The (3h)J(NCO2δ) coupling constant appears to be even smaller than the average value of backbone (3h)J(NC') couplings, consistent with more extensive local dynamics in protein side chains.",

keywords = "FKBP12, NMR detection, Sensitivity enhancement, Side chain-main chain hydrogen bonds",

author = "Aizhou Liu and Weidong Hu and Ananya Majumdar and Rosen, {Michael K.} and Patel, {Dinshaw J.}",

note = "Funding Information: This research was supported by NIH grant CA-46533 to D.J.P. and MCB-9817376 to M.K.R. We thank Dr. L.T. Kakalis for preparing13C/15N-labeled human FKBP12.",

year = "2000",

doi = "10.1023/A:1008373501591",

language = "English (US)",

volume = "17",

pages = "79--82",

journal = "Journal of biomolecular NMR",

issn = "0925-2738",

publisher = "Springer Netherlands",

number = "1",

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TY - JOUR

T1 - Detection of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins by sensitivity- enhanced NMR spectroscopy

AU - Liu, Aizhou

AU - Hu, Weidong

AU - Majumdar, Ananya

AU - Rosen, Michael K.

AU - Patel, Dinshaw J.

N1 - Funding Information: This research was supported by NIH grant CA-46533 to D.J.P. and MCB-9817376 to M.K.R. We thank Dr. L.T. Kakalis for preparing13C/15N-labeled human FKBP12.

PY - 2000

Y1 - 2000

N2 - We describe the direct observation of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the hydrogen acceptor side chain carboxylate carbon 13CO2(δ) of glutamate 54 and the hydrogen donor backbone amide 15N of methionine 49 in a 12 kDa protein, human FKBP12, is detected via the trans- hydrogen bond (3h)J(NCO2δ) coupling by employing a novel sensitivity- enhanced HNCO-type experiment, CPD-HNCO. The (3h)J(NCO2δ) coupling constant appears to be even smaller than the average value of backbone (3h)J(NC') couplings, consistent with more extensive local dynamics in protein side chains.

AB - We describe the direct observation of very weak side chain-main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the hydrogen acceptor side chain carboxylate carbon 13CO2(δ) of glutamate 54 and the hydrogen donor backbone amide 15N of methionine 49 in a 12 kDa protein, human FKBP12, is detected via the trans- hydrogen bond (3h)J(NCO2δ) coupling by employing a novel sensitivity- enhanced HNCO-type experiment, CPD-HNCO. The (3h)J(NCO2δ) coupling constant appears to be even smaller than the average value of backbone (3h)J(NC') couplings, consistent with more extensive local dynamics in protein side chains.

KW - FKBP12

KW - NMR detection

KW - Sensitivity enhancement

KW - Side chain-main chain hydrogen bonds

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