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Role of Cholesterol and its Biosynthetic Precursors on Membrane Organization and Dynamics: A Fluorescence Approach

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Abstract

Cholesterol is the most representative sterol present in membranes of higher eukaryotes, and is the end product of a long and multistep biosynthetic pathway. Lathosterol and zymosterol are biosynthetic precursors of cholesterol in Kandutsch-Russell and Bloch pathways, respectively. Lathosterol differs with cholesterol merely in the position of the double bond in the sterol ring, whereas zymosterol differs with cholesterol in position and number of double bonds. In this work, we have monitored the effect of cholesterol and its biosynthetic precursors (lathosterol and zymosterol) on membrane organization and dynamics in fluid and gel phase membranes. Toward this goal, we have utilized two fluorescent membrane probes, DPH and its cationic derivative TMA-DPH. Our results using these probes show that cholesterol and its biosynthetic precursors (lathosterol and zymosterol) exhibit similar trend in maintaining membrane organization and dynamics (as reported by fluorescence anisotropy and apparent rotational correlation time), in fluid phase POPC membranes. Notably, although lathosterol and zymosterol show similar trend in maintaining membrane organization and dynamics, the corresponding change for cholesterol is different in gel phase DPPC membranes. These results demonstrate that the position and number of double bonds in sterols is an important determinant in maintaining membrane physical properties. Our results assume significance since accumulation of precursors of cholesterol have been reported to be associated with severe pathological conditions.

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Fig. 1
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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

CDPX2:

X-linked chondrodysplasia punctata 2

DMPC:

1,2-dimyristoyl-sn-glycero-3-phosphocholine

DPH:

1,6-diphenyl-1,3,5-hexatriene

DPPC:

1,2-dipalmitoyl-sn-glycero-3- phosphocholine

LUV:

large unilamellar vesicle

POPC:

1-palmitoyl-2-oleoyl-sn-glycero-3- phosphocholine

TMA-DPH:

1-[4-(trimethylammonio)phenyl]-6-phenyl-1,3,5-hexatriene

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Acknowledgements

A.C. gratefully acknowledges support from CSIR Bhatnagar Fellowship. We gratefully acknowledge members of the Chattopadhyay laboratory and Dr. Parijat Sarkar (Stanford University) for their comments and suggestions.

Funding

CSIR Bhatnagar Fellowship, R90807

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Author notes

  1. Yamuna Devi Paila

    Present address: Moderna Inc, Cambridge, MA, 02139, USA

Authors and Affiliations

  1. CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007, India

    Sandeep Shrivastava, Yamuna Devi Paila & Amitabha Chattopadhyay

  2. Academy of Scientific and Innovative Research, Ghaziabad, 201 002, India

    Amitabha Chattopadhyay

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  1. Sandeep Shrivastava
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  2. Yamuna Devi Paila
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Contributions

AC, YDP and SS: conceptualized the project and designed experiments. SS and YDP: performed experiments. SS and YDP: analyzed the data. SS and AC: wrote the manuscript. AC: edited the manuscript, organized access to research facilities and funding, and provided overall supervision and mentoring.

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Correspondence to Amitabha Chattopadhyay.

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Shrivastava, S., Paila, Y.D. & Chattopadhyay, A. Role of Cholesterol and its Biosynthetic Precursors on Membrane Organization and Dynamics: A Fluorescence Approach. J Membrane Biol 256, 189–197 (2023). https://doi.org/10.1007/s00232-023-00278-w

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