Native Mass Spectrometry Analysis of Oligomerization States of Fluorescence Recovery Protein and Orange Carotenoid Protein: Two Proteins Involved in the Cyanobacterial Photoprotection Cycle

Overview

Journal Biochemistry

Specialty Biochemistry

Date 2016 Dec 21

PMID 27997134

Citations 14

Authors

Yue Lu

Haijun Liu

Rafael G Saer

Hao Zhang

Christine M Meyer

Veronica L Li

Liuqing Shi

Jeremy D King

Michael L Gross

Robert E Blankenship

Affiliations

Soon will be listed here.

Abstract

The orange carotenoid protein (OCP) and fluorescence recovery protein (FRP) are present in many cyanobacteria and regulate an essential photoprotection cycle in an antagonistic manner as a function of light intensity. We characterized the oligomerization states of OCP and FRP by using native mass spectrometry, a technique that has the capability of studying native proteins under a wide range of protein concentrations and molecular masses. We found that dimeric FRP is the predominant state at protein concentrations ranging from 3 to 180 μM and that higher-order oligomers gradually form at protein concentrations above this range. The OCP, however, demonstrates significantly different oligomerization behavior. Monomeric OCP (mOCP) dominates at low protein concentrations, with an observable population of dimeric OCP (dOCP). The ratio of dOCP to mOCP, however, increases proportionally with protein concentration. Higher-order OCP oligomers form at protein concentrations beyond 10 μM. Additionally, native mass spectrometry coupled with ion mobility allowed us to measure protein collisional cross sections and interrogate the unfolding of different FRP and OCP oligomers. We found that monomeric FRP exhibits a one-stage unfolding process, which could be correlated with its C-terminal bent crystal structure. The structural domain compositions of FRP and OCP are compared and discussed.

Citing Articles

Designed Cell-Penetrating Peptide Self-Assembly Featuring Distinctive Tertiary Structure.

Park J, Yamashita E, Yu J, Lee S, Hyun S ACS Omega. 2024; 9(30):32991-32999.

PMID: 39100342 PMC: 11292830. DOI: 10.1021/acsomega.4c04004.

Fuzzy Drug Targets: Disordered Proteins in the Drug-Discovery Realm.

Saurabh S, Nadendla K, Purohit S, Sivakumar P, Cetinel S ACS Omega. 2023; 8(11):9729-9747.

PMID: 36969402 PMC: 10034788. DOI: 10.1021/acsomega.2c07708.

The increasing role of structural proteomics in cyanobacteria.

Sound J, Bellamy-Carter J, Leney A Essays Biochem. 2022; 67(2):269-282.

PMID: 36503929 PMC: 10070481. DOI: 10.1042/EBC20220095.

Hydrodynamic conditions affect the proteomic profile of marine biofilms formed by filamentous cyanobacterium.

Romeu M, Dominguez-Perez D, Almeida D, Morais J, Araujo M, Osorio H NPJ Biofilms Microbiomes. 2022; 8(1):80.

PMID: 36253388 PMC: 9576798. DOI: 10.1038/s41522-022-00340-w.

Oligomerization processes limit photoactivation and recovery of the orange carotenoid protein.

Andreeva E, Nizinski S, Wilson A, Levantino M, De Zitter E, Munro R Biophys J. 2022; 121(15):2849-2872.

PMID: 35794830 PMC: 9388578. DOI: 10.1016/j.bpj.2022.07.004.

References

Brange J, Andersen L, Laursen E, Meyn G, Rasmussen E . Toward understanding insulin fibrillation. J Pharm Sci. 1997; 86(5):517-25. DOI: 10.1021/js960297s. View

Nettleton E, Tito P, Sunde M, Bouchard M, Dobson C, Robinson C . Characterization of the oligomeric states of insulin in self-assembly and amyloid fibril formation by mass spectrometry. Biophys J. 2000; 79(2):1053-65. PMC: 1301001. DOI: 10.1016/S0006-3495(00)76359-4. View

Lanucara F, Holman S, Gray C, Eyers C . The power of ion mobility-mass spectrometry for structural characterization and the study of conformational dynamics. Nat Chem. 2014; 6(4):281-94. DOI: 10.1038/nchem.1889. View

Ruotolo B, Benesch J, Sandercock A, Hyung S, Robinson C . Ion mobility-mass spectrometry analysis of large protein complexes. Nat Protoc. 2008; 3(7):1139-52. DOI: 10.1038/nprot.2008.78. View

Heck A, van den Heuvel R . Investigation of intact protein complexes by mass spectrometry. Mass Spectrom Rev. 2004; 23(5):368-89. DOI: 10.1002/mas.10081. View

Sutter M, Wilson A, Leverenz R, Lopez-Igual R, Thurotte A, Salmeen A . Crystal structure of the FRP and identification of the active site for modulation of OCP-mediated photoprotection in cyanobacteria. Proc Natl Acad Sci U S A. 2013; 110(24):10022-7. PMC: 3683793. DOI: 10.1073/pnas.1303673110. View

Fitzgerald M, Chernushevich I, Standing K, Whitman C, Kent S . Probing the oligomeric structure of an enzyme by electrospray ionization time-of-flight mass spectrometry. Proc Natl Acad Sci U S A. 1996; 93(14):6851-6. PMC: 38896. DOI: 10.1073/pnas.93.14.6851. View

Marcoux J, Robinson C . Twenty years of gas phase structural biology. Structure. 2013; 21(9):1541-50. DOI: 10.1016/j.str.2013.08.002. View

Gupta S, Guttman M, Leverenz R, Zhumadilova K, Pawlowski E, Petzold C . Local and global structural drivers for the photoactivation of the orange carotenoid protein. Proc Natl Acad Sci U S A. 2015; 112(41):E5567-74. PMC: 4611662. DOI: 10.1073/pnas.1512240112. View

10.

Wilson A, Kinney J, Zwart P, Punginelli C, DHaene S, Perreau F . Structural determinants underlying photoprotection in the photoactive orange carotenoid protein of cyanobacteria. J Biol Chem. 2010; 285(24):18364-75. PMC: 2881762. DOI: 10.1074/jbc.M110.115709. View

11.

Wilson A, Punginelli C, Gall A, Bonetti C, Alexandre M, Routaboul J . A photoactive carotenoid protein acting as light intensity sensor. Proc Natl Acad Sci U S A. 2008; 105(33):12075-80. PMC: 2575289. DOI: 10.1073/pnas.0804636105. View

12.

Ruotolo B, Hyung S, Robinson P, Giles K, Bateman R, Robinson C . Ion mobility-mass spectrometry reveals long-lived, unfolded intermediates in the dissociation of protein complexes. Angew Chem Int Ed Engl. 2007; 46(42):8001-4. DOI: 10.1002/anie.200702161. View

13.

Hall Z, Politis A, Robinson C . Structural modeling of heteromeric protein complexes from disassembly pathways and ion mobility-mass spectrometry. Structure. 2012; 20(9):1596-609. DOI: 10.1016/j.str.2012.07.001. View

14.

Zhang H, Cui W, Gross M, Blankenship R . Native mass spectrometry of photosynthetic pigment-protein complexes. FEBS Lett. 2013; 587(8):1012-20. PMC: 3856239. DOI: 10.1016/j.febslet.2013.01.005. View

15.

Hall Z, Politis A, Bush M, Smith L, Robinson C . Charge-state dependent compaction and dissociation of protein complexes: insights from ion mobility and molecular dynamics. J Am Chem Soc. 2012; 134(7):3429-38. DOI: 10.1021/ja2096859. View

16.

Hong P, Koza S, Bouvier E . Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and their Aggregates. J Liq Chromatogr Relat Technol. 2013; 35(20):2923-2950. PMC: 3556795. DOI: 10.1080/10826076.2012.743724. View

17.

Ali M, Imperiali B . Protein oligomerization: how and why. Bioorg Med Chem. 2005; 13(17):5013-20. DOI: 10.1016/j.bmc.2005.05.037. View

18.

Benesch J, Ruotolo B . Mass spectrometry: come of age for structural and dynamical biology. Curr Opin Struct Biol. 2011; 21(5):641-9. PMC: 3193349. DOI: 10.1016/j.sbi.2011.08.002. View

19.

Nishiyama Y, Allakhverdiev S, Murata N . A new paradigm for the action of reactive oxygen species in the photoinhibition of photosystem II. Biochim Biophys Acta. 2006; 1757(7):742-9. DOI: 10.1016/j.bbabio.2006.05.013. View

20.

Goodsell D, Olson A . Structural symmetry and protein function. Annu Rev Biophys Biomol Struct. 2000; 29:105-53. DOI: 10.1146/annurev.biophys.29.1.105. View