Evidence map›Paper›PMID 39248532›Full record

ArticleJournal of visualized experiments : JoVE2024

MultiBac System-based Purification and Biophysical Characterization of Human Myosin-7a.

Marvin Wright, Shayna Redford, Jacob Vehar, Kevin C Courtney, Neil Billington, Rong Liu

Abstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Marvin Wright *Department of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University.
Shayna Redford *Department of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University.
Jacob VeharDepartment of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University.
Kevin C CourtneyDepartment of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University.
Neil BillingtonDepartment of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University; Microscope Imaging Facility, West Virginia University.
Rong LiuDepartment of Biochemistry & Molecular Medicine, School of Medicine, West Virginia University; rong.liu@hsc.wvu.edu.

Funding

WV INBRE: The Inhibitor of Growth Family Member 4 (ING4) inhibits L-Type Amino Acid Transporter 1 (LAT1) expression to suppress Breast CancerP20GM103434 · NIGMS · MARSHALL UNIVERSITY · PI GARY O RANKIN · 2012 to 2026
$61.1M
VS-CoBRE Administrative CoreP20GM144230 · NIGMS · WEST VIRGINIA UNIVERSITY · PI Visvanathan Ramamurthy · 2022 to 2026
$13.9M
NIGMS NIH HHS P20 GM103434NIGMS NIH HHS P20 GM144230
6 · The paper itself

Abstract

Myosin-7a is an actin-based motor protein vital for auditory and visual processes. Mutations in myosin-7a lead to Usher syndrome type 1, the most common and severe form of deaf-blindness in humans. It is hypothesized that myosin-7a forms a transmembrane adhesion complex with other Usher proteins, essential for the structural-functional integrity of photoreceptor and cochlear hair cells. However, due to the challenges in obtaining pure, intact protein, the exact functional mechanisms of human myosin-7a remain elusive, with limited structural and biomechanical studies available. Recent studies have shown that mammalian myosin-7a is a multimeric motor complex consisting of a heavy chain and three types of light chains: regulatory light chain (RLC), calmodulin, and calmodulin-like protein 4 (CALML4). Unlike calmodulin, CALML4 does not bind to calcium ions. Both the calcium-sensitive, and insensitive calmodulins are critical for mammalian myosin-7a for proper fine-tuning of its mechanical properties. Here, we describe a detailed method to produce recombinant human myosin-7a holoenzyme using the MultiBac Baculovirus protein expression system. This yields milligram quantities of high-purity full-length protein, allowing for its biochemical and biophysical characterization. We further present a protocol for assessing its mechanical and motile properties using tailored in vitro motility assays and fluorescence microscopy. The availability of the intact human myosin-7a protein, along with the detailed functional characterization protocol described here, paves the way for further investigations into the molecular aspects of myosin-7a in vision and hearing.

Indexed as

Myosin VIIaAnimalsBaculoviridaeHumansMyosinsSf9 CellsSpodopteraMYO7A protein, humanMyosinsMyosin VIIa

Identifiers

PMID39248532
PMCPMC11633084

What Socratic holds

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Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.