effect of palmitoylation on claudin-14 tight junction proteinsfundamentals of biochemistry: life at...

Laurie Muok, Kathryn Piston, Nandhini Rajagopal, and Dr. Shikha Nangia Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse NY 13244, United States Effect of Palmitoylation on Claudin-14 Tight Junction Proteins 1 1 item selected Introduction Results Acknowledgements Methods Conclusions References Claudin-14 Palmitoylated Claudin-14 Coarse Grain models of claudin-14 were generated with and without palmitoylation chains The atomistic structures used to create the coarse grain models using Martini v2.2 force field The systems were solvated with standard Martini CG water with 0.15 M NaCl All CG MD simulations were performed in GROMACS Simulations were run in triplicates Each run was performed for 10 μs Claudins are membrane proteins that form paracellular barriers and pores that determine tight junction permeability. The claudin family has 27 known claudin proteins in humans. The expression levels of the claudins are tissue specific. Claudin-14 is found in the inner ear and the kidneys. Defects in claudin-14 expression can lead to: deafness, kidney stones, and reduced bone density Previous study showed that specific mutations of palmitoylation sites in claudin-14 affect its ability to create tight junctions Palmitoylation of claudin- 14 protein impacts its interactions with the surrounding lipids Computational resources were provided by Information and Technology Services at Syracuse University (Eric Sedore, Larne Pekowsky, and Michael R. Brady) Palmitoylation of Claudin-5 Proteins Influences Their Lipid Domain Affinity and Tight Junction Assembly at the Blood–Brain Barrier Interface. N. Rajagopal, F. J. Irudayanathan, S. Nangia, Journal of Physical Chemistry B. 123, 983-993 (2019). CLDN14 claudin 14 [Homo sapiens (human)] - Gene - NCBI. (2019, June 3). Retrieved from https://www.ncbi.nlm.nih.gov/gene/23562 Genetics Home Reference. (2019, June 11). What are proteins and what do they do?. Retrieved from https://ghr.nlm.nih.gov/primer/howgeneswo rk/protein Voet, D., Voet, J. G., & Pratt, C. W. (2011). Lipids and Biological Membranes. In Fundamentals of Biochemistry: Life at the Molecular Level, 4th Edition: Life at the Molecular Level(4th ed., pp. 241-290). MASTAVQLLGFLLSFLGMVGTLITTILPHWRRTAHVGTNILTAVSYLKGLWMECVWHSTGIYQCQIYRSLLALPQDLQAARALMVISCLLSGIACACAVIGMKCTRCAKGTPAKTTFAILGGTLFILAGLLCMVAVSWTTNDVVQNFYNPLLPSGMKFEIGQALYLGFISSSLSLIGGTLLCLSCQDEAPYRPYQAPPRATTTTANTAPAYQPPAAYKDNRAPSVTSATHSGYRLNDYV The simulations showed that when palmitoylated, claudin 14’s interactions with the DPPC increased. This was also observed in six other claudins. These results can lead to further studies to see how two claudins interact with each other when they are in the same membrane. DPPC Lower Leaflet Palm DPPC Lower Leaflet Non Palm 0 1 2 3 4 Time (µs) Normalized Contacts DOPC DPPC CHOL 0 1 2 3 4 Time (µs) Normalized Contacts DOPC DPPC CHOL DOPC Lower Leaflet Non Palm DOPC Lower Leaflet Palm X-axis X-axis X-axis X-axis Y-axis Y-axis Y-axis Y-axis Non Palmitoylated Palmitoylated

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Laurie Muok, Kathryn Piston, Nandhini Rajagopal, and Dr. Shikha Nangia Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse NY 13244, United States

Effect of Palmitoylation on Claudin-14 Tight Junction Proteins

11 item selected

Introduction

Results

Acknowledgements

Methods

Conclusions

ReferencesClaudin-14 Palmitoylated Claudin-14Coarse Grain models of claudin-14 were generated with and without palmitoylation chains

The atomistic structures used to create the coarse grain models using Martini v2.2 force field

The systems were solvated with standard Martini CG water with 0.15 M NaCl

All CG MD simulations were performed in GROMACS

Simulations were run in triplicates

Each run was performed for 10 μs

Claudins are membrane proteins that form paracellular barriers and pores that determine tight junction permeability. The claudin family has 27 known claudin proteins in humans. The expression levels of the claudins are tissue specific. Claudin-14 is found in the inner ear and the kidneys. Defects in claudin-14 expression can lead to: deafness, kidney stones, and reduced bone density

Previous study showed that specific mutations of palmitoylation sites in claudin-14 affect its ability to create tight junctions

Palmitoylation of claudin-14 protein impacts its interactions with the surrounding lipids

Computational resources were provided by Information and Technology Services

at Syracuse University (Eric Sedore, Larne Pekowsky, and Michael R. Brady)

Palmitoylation of Claudin-5 Proteins Influences Their Lipid Domain Affinity and

Tight Junction Assembly at the Blood–Brain Barrier Interface. N. Rajagopal, F. J.

Irudayanathan, S. Nangia, Journal of Physical Chemistry B. 123, 983-993 (2019).

CLDN14 claudin 14 [Homo sapiens (human)] -Gene - NCBI. (2019, June 3). Retrieved from https://www.ncbi.nlm.nih.gov/gene/23562Genetics Home Reference. (2019, June 11).

What are proteins and what do they do?. Retrieved from

https://ghr.nlm.nih.gov/primer/howgeneswork/protein

Voet, D., Voet, J. G., & Pratt, C. W. (2011). Lipids and Biological Membranes. In

Fundamentals of Biochemistry: Life at the Molecular Level, 4th Edition: Life at the

Molecular Level(4th ed., pp. 241-290).

MASTAVQLLGFLLSFLGMVGTLITTILPHWRRTAHVGTNILTAVSYLKGLWMECVWHSTGIYQCQIYRSLLALPQDLQAARALMVISCLLSGIACACAVIGMKCTRCAKGTPAKTTFAILGGTLFILAGLLCMVAVSWTTNDVVQNFYNPLLPSGMKFEIGQALYLGFISSSLSLIGGTLLCLSCQDEAPYRPYQAPPRATTTTANTAPAYQPPAAYKDNRAPSVTSATHSGYRLNDYV

The simulations showed that when palmitoylated, claudin 14’s interactions with the DPPC increased. This was also observed in six other claudins. These results can lead to

further studies to see how two claudins interact with each other when they are in the same membrane.

DPPC Lower Leaflet PalmDPPC Lower Leaflet Non Palm

0 1 2 3 4Time (µs)

Nor

mal

ized

Con

tact

s DOPCDPPCCHOL

0 1 2 3 4Time (µs)

Nor

mal