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Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry

Publication ,  Journal Article
Schwartz, BL; Gale, DC; Smith, RD; Chilkoti, A; Stayton, PS
Published in: Journal of Mass Spectrometry
January 1, 1995

The relative non‐covalent binding behavior of two small molecule ligands to the tetrameric protein streptavidin was investigated by electrospray ionization mass spectrometry (ESI‐MS). An extended m/z range quadrupole mass spectrometer was employed to observe the intact multimeric form of the protein and to probe the relative stabilities of the tetrameric protein‐ ligand complexes in the gas phase. Various protein:ligand molar ratio concentrations and incubation times were studied for the ligands biotin (Kd ≈ 10−15 M) and iminobiotin (Kd ≈ 10−7 M), in combination with the adjustment of variables in the ESI atmosphere‐vacuum interface region. Positive‐ion ESI‐MS of a 1:7 molar ratio streptvidin–biotin sample produced peaks corresponding to the intact tetrameric complex (16+ to 14+ charge states) with four ligands binding to the active from of the protein. Only the expected specific non‐covalent complexes of four ligand molecules binding to the protein tetramer were detected, consistent with known solution behavior and without the appearance of any random aggregation. However, under identical interface conditions, complete iminobiotin binding to the protein was not observed, even when higher ligand concentrations, longer complexation times and gentler interface conditions were employed. The thermally induced dissociation (TID) of the non‐covalent complexes was studied by applying more severe conditions in the ESI interface region. In addition, observation of an intact streptavidin–biotinylated oligonucleotide non‐covalent complex (17– to 14– tetrameric charge states) by negative‐ion ESI‐MS at m/z 4500–5000 is presented. The results show that the complexes observed in the mass spectrometer are representative of those that exist in solution, under the ESI conditions employed, and that the ability to observe such complexes is at least qualitatively related to stability in solution. Copyright © 1995 John Wiley & Sons, Ltd.

Duke Scholars

Published In

Journal of Mass Spectrometry

DOI

EISSN

1096-9888

ISSN

1076-5174

Publication Date

January 1, 1995

Volume

30

Issue

8

Start / End Page

1095 / 1102

Related Subject Headings

  • Analytical Chemistry
  • 34 Chemical sciences
  • 03 Chemical Sciences
 

Citation

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ICMJE
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Schwartz, B. L., Gale, D. C., Smith, R. D., Chilkoti, A., & Stayton, P. S. (1995). Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry. Journal of Mass Spectrometry, 30(8), 1095–1102. https://doi.org/10.1002/jms.1190300806
Schwartz, B. L., D. C. Gale, R. D. Smith, A. Chilkoti, and P. S. Stayton. “Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry.” Journal of Mass Spectrometry 30, no. 8 (January 1, 1995): 1095–1102. https://doi.org/10.1002/jms.1190300806.
Schwartz BL, Gale DC, Smith RD, Chilkoti A, Stayton PS. Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry. Journal of Mass Spectrometry. 1995 Jan 1;30(8):1095–102.
Schwartz, B. L., et al. “Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry.” Journal of Mass Spectrometry, vol. 30, no. 8, Jan. 1995, pp. 1095–102. Scopus, doi:10.1002/jms.1190300806.
Schwartz BL, Gale DC, Smith RD, Chilkoti A, Stayton PS. Investigation of non‐covalent ligand binding to the intact streptavidin tetramer by electrospray ionization mass spectrometry. Journal of Mass Spectrometry. 1995 Jan 1;30(8):1095–1102.
Journal cover image

Published In

Journal of Mass Spectrometry

DOI

EISSN

1096-9888

ISSN

1076-5174

Publication Date

January 1, 1995

Volume

30

Issue

8

Start / End Page

1095 / 1102

Related Subject Headings

  • Analytical Chemistry
  • 34 Chemical sciences
  • 03 Chemical Sciences