Dataset Information

Cooperative polarization of MCAM/CD146 and ERM family proteins in melanoma

ABSTRACT: The WRAMP structure is a protein network associated with tail-end actomyosin contractility, membrane retraction, and directional persistence during cell migration. A marker of WRAMP structures is melanoma cell adhesion molecule (MCAM) which dynamically polarizes to the cell rear. However, factors that mediate MCAM polarization are still unknown. In this study, BioID using MCAM as bait identifies the ERM family proteins, moesin, ezrin, and radixin, as WRAMP structure components. We also present a novel image analysis pipeline, Protein Polarity by Percentile (“3P”), which classifies protein polarization using machine learning and facilitates quantitative analysis. Using 3P, we find that depletion of moesin, and to a lesser extent ezrin, decreases the proportion of cells with polarized MCAM. Furthermore, although co-polarized MCAM and ERM proteins show high spatial overlap, 3P identifies subpopulations with ERM proteins closer to the cell periphery. Live-cell imaging confirms that MCAM and ERM protein polarization is tightly coordinated, but ERM proteins enrich at the cell edge first. Finally, deletion of a juxtamembrane segment in MCAM previously shown to promote ERM protein interactions impedes MCAM polarization. Our findings highlight the requirement for ERM proteins in recruitment of MCAM to WRAMP structures and an advanced computational tool to characterize protein polarization.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Melanocyte, Cell Culture

SUBMITTER: Christopher Ebmeier

LAB HEAD: Natalie G Ahn

PROVIDER: PXD047722 | Pride | 2024-05-22

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	CCE20211015_Suzannah_Miller_BioID_constructs.fasta	Fasta
	CCE20220117_Suzannah_Miller_AhnLab_s13_No_Construct_DMSO_C_hfx1.raw	Raw
	CCE20220117_Suzannah_Miller_AhnLab_s13_No_Construct_DMSO_C_hfx2.raw	Raw
	CCE20220117_Suzannah_Miller_AhnLab_s3_No_Construct_DMSO_A_hfx1.raw	Raw
	CCE20220117_Suzannah_Miller_AhnLab_s3_No_Construct_DMSO_A_hfx2.raw	Raw

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Publications

Cooperative polarization of MCAM/CD146 and ERM family proteins in melanoma.

Miller Suzannah G SG Hoh Maria M Ebmeier Christopher C CC Tay Jian Wei JW Ahn Natalie G NG

Molecular biology of the cell 20231220 3

The WRAMP structure is a protein network associated with tail-end actomyosin contractility, membrane retraction, and directional persistence during cell migration. A marker of WRAMP structures is melanoma cell adhesion molecule (MCAM) which dynamically polarizes to the cell rear. However, factors that mediate MCAM polarization are still unknown. In this study, BioID using MCAM as bait identifies the ERM family proteins, moesin, ezrin, and radixin, as WRAMP structure components. We also present a ...[more]

PMID: 38117590

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