Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements

Helle Johansson; Malene Ringkjøbing Jensen; Henrik Gesmar; Sebastian Meier; Joachim Møllesøe Vinther; Camille Keeler; Michael E. Hodsdon; Jens J. Led

doi:10.1021/ja503546j

Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements

Helle Johansson, Malene Ringkjøbing Jensen, Henrik Gesmar, Sebastian Meier, Joachim Møllesøe Vinther, Camille Keeler, Michael E. Hodsdon, Jens J. Led

23 Citationer (Scopus)

Abstract

Weak and transient protein-protein interactions underlie numerous biological processes. However, the location of the interaction sites of the specific complexes and the effect of transient, nonspecific protein-protein interactions often remain elusive. We have investigated the weak self-association of human growth hormone (hGH, K_D = 0.90 ± 0.03 mM) at neutral pH by the paramagnetic relaxation enhancement (PRE) of the amide protons induced by the soluble paramagnetic relaxation agent, gadodiamide (Gd(DTPA-BMA)). Primarily, it was found that the PREs are in agreement with the general Hwang-Freed model for relaxation by translational diffusion (J. Chem. Phys. 1975, 63, 4017-4025), only if crowding effects on the diffusion in the protein solution are taken into account. Second, by measuring the PREs of the amide protons at increasing hGH concentrations and a constant concentration of the relaxation agent, it is shown that a distinction can be made between residues that are affected only by transient, nonspecific protein-protein interactions and residues that are involved in specific protein-protein associations. Thus, the PREs of the former residues increase linearly with the hGH concentration in the entire concentration range because of a reduction of the diffusion caused by the transient, nonspecific protein-protein interactions, while the PREs of the latter residues increase only at the lower hGH concentrations but decrease at the higher concentrations because of specific protein-protein associations that impede the access of gadodiamide to the residues of the interaction surface. finally, it is found that the ultraweak aggregation of hGH involves several interaction sites that are located in patches covering a large part of the protein surface.

Originalsprog	Engelsk
Tidsskrift	Journal of the American Chemical Society
Vol/bind	136
Udgave nummer	29
Sider (fra-til)	10277-10286
Antal sider	10
ISSN	0002-7863
DOI	https://doi.org/10.1021/ja503546j
Status	Udgivet - 23 jul. 2014

Emneord

Det Natur- og Biovidenskabelige Fakultet

Adgang til dokumentet

10.1021/ja503546j

Citationsformater

Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements. / Johansson, Helle; Jensen, Malene Ringkjøbing; Gesmar, Henrik et al.

I: Journal of the American Chemical Society, Bind 136, Nr. 29, 23.07.2014, s. 10277-10286.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

@article{76005245ab0e408489ec5cc7ff1345e1,

title = "Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements",

abstract = "Weak and transient protein-protein interactions underlie numerous biological processes. However, the location of the interaction sites of the specific complexes and the effect of transient, nonspecific protein-protein interactions often remain elusive. We have investigated the weak self-association of human growth hormone (hGH, KD = 0.90 ± 0.03 mM) at neutral pH by the paramagnetic relaxation enhancement (PRE) of the amide protons induced by the soluble paramagnetic relaxation agent, gadodiamide (Gd(DTPA-BMA)). Primarily, it was found that the PREs are in agreement with the general Hwang-Freed model for relaxation by translational diffusion (J. Chem. Phys. 1975, 63, 4017-4025), only if crowding effects on the diffusion in the protein solution are taken into account. Second, by measuring the PREs of the amide protons at increasing hGH concentrations and a constant concentration of the relaxation agent, it is shown that a distinction can be made between residues that are affected only by transient, nonspecific protein-protein interactions and residues that are involved in specific protein-protein associations. Thus, the PREs of the former residues increase linearly with the hGH concentration in the entire concentration range because of a reduction of the diffusion caused by the transient, nonspecific protein-protein interactions, while the PREs of the latter residues increase only at the lower hGH concentrations but decrease at the higher concentrations because of specific protein-protein associations that impede the access of gadodiamide to the residues of the interaction surface. finally, it is found that the ultraweak aggregation of hGH involves several interaction sites that are located in patches covering a large part of the protein surface.",

keywords = "Faculty of Science",

author = "Helle Johansson and Jensen, {Malene Ringkj{\o}bing} and Henrik Gesmar and Sebastian Meier and Vinther, {Joachim M{\o}lles{\o}e} and Camille Keeler and Hodsdon, {Michael E.} and Led, {Jens J.}",

year = "2014",

month = jul,

day = "23",

doi = "10.1021/ja503546j",

language = "English",

volume = "136",

pages = "10277--10286",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "ACS Publications",

number = "29",

}

TY - JOUR

T1 - Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements

AU - Johansson, Helle

AU - Jensen, Malene Ringkjøbing

AU - Gesmar, Henrik

AU - Meier, Sebastian

AU - Vinther, Joachim Møllesøe

AU - Keeler, Camille

AU - Hodsdon, Michael E.

AU - Led, Jens J.

PY - 2014/7/23

Y1 - 2014/7/23

N2 - Weak and transient protein-protein interactions underlie numerous biological processes. However, the location of the interaction sites of the specific complexes and the effect of transient, nonspecific protein-protein interactions often remain elusive. We have investigated the weak self-association of human growth hormone (hGH, KD = 0.90 ± 0.03 mM) at neutral pH by the paramagnetic relaxation enhancement (PRE) of the amide protons induced by the soluble paramagnetic relaxation agent, gadodiamide (Gd(DTPA-BMA)). Primarily, it was found that the PREs are in agreement with the general Hwang-Freed model for relaxation by translational diffusion (J. Chem. Phys. 1975, 63, 4017-4025), only if crowding effects on the diffusion in the protein solution are taken into account. Second, by measuring the PREs of the amide protons at increasing hGH concentrations and a constant concentration of the relaxation agent, it is shown that a distinction can be made between residues that are affected only by transient, nonspecific protein-protein interactions and residues that are involved in specific protein-protein associations. Thus, the PREs of the former residues increase linearly with the hGH concentration in the entire concentration range because of a reduction of the diffusion caused by the transient, nonspecific protein-protein interactions, while the PREs of the latter residues increase only at the lower hGH concentrations but decrease at the higher concentrations because of specific protein-protein associations that impede the access of gadodiamide to the residues of the interaction surface. finally, it is found that the ultraweak aggregation of hGH involves several interaction sites that are located in patches covering a large part of the protein surface.

AB - Weak and transient protein-protein interactions underlie numerous biological processes. However, the location of the interaction sites of the specific complexes and the effect of transient, nonspecific protein-protein interactions often remain elusive. We have investigated the weak self-association of human growth hormone (hGH, KD = 0.90 ± 0.03 mM) at neutral pH by the paramagnetic relaxation enhancement (PRE) of the amide protons induced by the soluble paramagnetic relaxation agent, gadodiamide (Gd(DTPA-BMA)). Primarily, it was found that the PREs are in agreement with the general Hwang-Freed model for relaxation by translational diffusion (J. Chem. Phys. 1975, 63, 4017-4025), only if crowding effects on the diffusion in the protein solution are taken into account. Second, by measuring the PREs of the amide protons at increasing hGH concentrations and a constant concentration of the relaxation agent, it is shown that a distinction can be made between residues that are affected only by transient, nonspecific protein-protein interactions and residues that are involved in specific protein-protein associations. Thus, the PREs of the former residues increase linearly with the hGH concentration in the entire concentration range because of a reduction of the diffusion caused by the transient, nonspecific protein-protein interactions, while the PREs of the latter residues increase only at the lower hGH concentrations but decrease at the higher concentrations because of specific protein-protein associations that impede the access of gadodiamide to the residues of the interaction surface. finally, it is found that the ultraweak aggregation of hGH involves several interaction sites that are located in patches covering a large part of the protein surface.

KW - Faculty of Science

U2 - 10.1021/ja503546j

DO - 10.1021/ja503546j

M3 - Journal article

C2 - 24969589

SN - 0002-7863

VL - 136

SP - 10277

EP - 10286

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 29

ER -

Specific and nonspecific interactions in ultraweak protein−protein associations revealed by solvent paramagnetic relaxation enhancements

Abstract

Emneord

Adgang til dokumentet

Fingeraftryk

Citationsformater