Generic placeholder image

Current Protein & Peptide Science

Author(s): Ryan R. Manning, Ryan E. Holcomb, Derrick S. Katayama, Giovanni M. Pauletti, Shawn N. Grant, Jan S. Rosenbaum and Mark Cornell Manning*

DOI: 10.2174/1389203724666221102111145

DownloadDownload PDF Flyer Cite As
Characterization of Oligomer Formation of Surfactant Protein-D (SP-D) Using AF4-MALLS

Page: [862 - 873] Pages: 12

  • * (Excluding Mailing and Handling)

Abstract

Background: Surfactant protein-S (SP-D) is a naturally occurring lung protein with the potential to treat pulmonary infections. A recombinant surfactant protein-D (SP-D) has been produced and was previously found to exist in multiple oligomeric states.

Introduction: Separation and characterization of interconverting oligomeric states of a protein can be difficult using chromatographic methods, so an alternative separation technique was employed for SPD to characterize the different association states that exist.

Methods: Samples of SP-D were analyzed using asymmetrical flow field-flow fractionation (AF4) using UV and multi-angle laser light scattering (MALLS) detection. The AF4 method appears to be able to separate species as small as the monomer up to the dodecamer (the dominant species) to much larger species with a molar mass greater than 5 MDa.

Results: Consistent elution of four distinct peaks was observed after repeated injections. The largest species observed under the last peak (labeled as Peak 4) were termed “unstructured multimers” and were resolved fairly well from the other species. The AF4-MALLS data suggest that only a small fraction of Peak 4 truly corresponds to high molar mass unstructured multimers. All other peaks demonstrated significant molar mass homogeneity consistent with AFM results.

Conclusion: AF4-MALLS technology appears to be a powerful analytical approach to characterize the complex and dynamic interplay among different protein oligomeric species of SP-D in an aqueous solution.

Graphical Abstract

[1]
Crouch, E.; Persson, A.; Chang, D.; Heuser, J. Molecular structure of pulmonary surfactant protein D (SP-D). J. Biol. Chem., 1994, 269(25), 17311-17319.
[http://dx.doi.org/10.1016/S0021-9258(17)32556-5] [PMID: 8006040]
[2]
Sorensen, G.L.; Hoegh, S.V.; Leth-Larsen, R.; Thomsen, T.H.; Floridon, C.; Smith, K.; Kejling, K.; Tornoe, I.; Crouch, E.C.; Holmskov, U. Multimeric and trimeric subunit SP-D are interconvertible structures with distinct ligand interaction. Mol. Immunol., 2009, 46(15), 3060-3069.
[http://dx.doi.org/10.1016/j.molimm.2009.06.005] [PMID: 19577304]
[3]
Crouch, E.; Chang, D.; Rust, K.; Persson, A.; Heuser, J. Recombinant pulmonary surfactant protein D. Post-translational modification and molecular assembly. J. Biol. Chem., 1994, 269(22), 15808-15813.
[http://dx.doi.org/10.1016/S0021-9258(17)40752-6] [PMID: 8195236]
[4]
Arroyo, R.; Martín-González, A.; Echaide, M.; Jain, A.; Brondyk, W.H.; Rosenbaum, J.; Moreno-Herrero, F.; Pérez-Gil, J. Supramolecular assembly of human pulmonary surfactant protein SP-D. J. Mol. Biol., 2018, 430(10), 1495-1509.
[http://dx.doi.org/10.1016/j.jmb.2018.03.027] [PMID: 29626540]
[5]
Crouch, E.C. Surfactant protein-D and pulmonary host defense. Respir. Res., 2000, 1(2), 6.
[http://dx.doi.org/10.1186/rr19] [PMID: 11667972]
[6]
Håkansson, K.; Reid, K.B.M. Collectin structure: A review. Protein Sci., 2000, 9(9), 1607-1617.
[http://dx.doi.org/10.1110/ps.9.9.1607] [PMID: 11045608]
[7]
Crouch, E.; McDonald, B.; Smith, K.; Cafarella, T.; Seaton, B.; Head, J. Contributions of phenylalanine 335 to ligand recognition by hu-man surfactant protein D: Ring interactions with SP-D ligands. J. Biol. Chem., 2006, 281(26), 18008-18014.
[http://dx.doi.org/10.1074/jbc.M601749200] [PMID: 16636058]
[8]
Arroyo, R.; Echaide, M.; Moreno-Herrero, F.; Perez-Gil, J.; Kingma, P.S. Functional characterization of the different oligomeric forms of human surfactant protein SP-D. Biochim. Biophys. Acta, 2020, 1868(8), 140436.
[http://dx.doi.org/10.1016/j.bbapap.2020.140436]
[9]
Rambaldi, D.C.; Zattoni, A.; Reschiglian, P.; Colombo, R.; De Lorenzi, E. In vitro amyloid Aβ1-42 peptide aggregation monitoring by asymmetrical flow field-flow fractionation with multi-angle light scattering detection. Anal. Bioanal. Chem., 2009, 394(8), 2145-2149.
[http://dx.doi.org/10.1007/s00216-009-2899-1] [PMID: 19562325]
[10]
Hawe, A.; Romeijn, S.; Filipe, V.; Jiskoot, W. Asymmetrical flow field-flow fractionation method for the analysis of submicron protein aggregates. J. Pharm. Sci., 2012, 101(11), 4129-4139.
[http://dx.doi.org/10.1002/jps.23298] [PMID: 22911663]
[11]
Yohannes, G.; Jussila, M.; Hartonen, K.; Riekkola, M.L. Asymmetrical flow field-flow fractionation technique for separation and charac-terization of biopolymers and bioparticles. J. Chromatogr. A, 2011, 1218(27), 4104-4116.
[http://dx.doi.org/10.1016/j.chroma.2010.12.110] [PMID: 21292269]
[12]
Pavišić R.; Dodig, I.; Horvatić A.; Mijić L.; Sedić M.; Linarić M.R.; Sovulj, I.G.; Preočanin, T.; Krajačić M.B.; Cindrić M. Differences between reversible (self-association) and irreversible aggregation of rHuG-CSF in carbohydrate and polyol formulations. Eur. J. Pharm. Biopharm., 2010, 76(3), 357-365.
[http://dx.doi.org/10.1016/j.ejpb.2010.09.006] [PMID: 20854908]
[13]
Choi, J.; Lee, S.; Linares-Pastén, J.A.; Nilsson, L. Study on oligomerization of glutamate decarboxylase from Lactobacillus brevis using asymmetrical flow field-flow fractionation (AF4) with light scattering techniques. Anal. Bioanal. Chem., 2018, 410(2), 451-458.
[http://dx.doi.org/10.1007/s00216-017-0735-6] [PMID: 29167934]
[14]
Efrati, H.; Hawgood, S.; Williams, M.C.; Hong, K.; Benson, B.J. Divalent cation and hydrogen ion effects on the structure and surface activity of pulmonary surfactant. Biochemistry, 1987, 26(24), 7986-7993.
[http://dx.doi.org/10.1021/bi00398a066] [PMID: 3427118]
[15]
Duvoix, A.; Mackay, R.M.; Henderson, N.; McGreal, E.; Postle, A.; Reid, K.; Clark, H. Physiological concentration of calcium inhibits elastase-induced cleavage of a functional recombinant fragment of surfactant protein D. Immunobiology, 2011, 216(1-2), 72-79.
[http://dx.doi.org/10.1016/j.imbio.2010.03.006] [PMID: 20378199]
[16]
Crouch, E.C. Collectins and pulmonary host defense. Am. J. Respir. Cell Mol. Biol., 1998, 19(2), 177-201.
[http://dx.doi.org/10.1165/ajrcmb.19.2.140] [PMID: 9698590]