Molecular analysis of a secretory organelle: Structure and function of synaptic vesicle-specific proteins

SUMMARYNanobodies (nAbs) are small, minimal antibodies that have distinct attributes that make them uniquely suited for certain biomedical research, diagnostic and therapeutic applications. Prominent uses include as intracellular antibodies or intrabodies to bind and deliver cargo to specific proteins and/or subcellular sites within cells, and as nanoscale immunolabels for enhanced tissue penetration and improved spatial imaging resolution. Here, we report the generation and validation of nAbs against a set of proteins prominently expressed at specific subcellular sites in brain neurons. We describe a novel hierarchical validation pipeline to systematically evaluate nAbs isolated by phage display for effective and specific use as intrabodies and immunolabels in mammalian cells including brain neurons. These nAbs form part of a robust toolbox for targeting proteins with distinct and highly spatially-restricted subcellular localization in mammalian brain neurons, allowing for visualization and/or modulation of structure and function at those sites.

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Role of Lung Surfactant-Specific Proteins in Surfactant Structure and Function

Current Concepts in Surfactant Research - Progress in Respiratory Research ◽

10.1159/000410001 ◽

2015 ◽

pp. 83-92 ◽

Cited By ~ 10

Author(s):

Bradley J. Benson ◽

Mary C. Williams ◽

Samuel Hawgood ◽

Tania Sargeant

Keyword(s):

Lung Surfactant ◽

Structure And Function ◽

Specific Proteins ◽

And Function

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Gap junction Heterogeneity in Liver, Heart, and Lens

Physiology ◽

10.1152/physiologyonline.1988.3.5.206 ◽

1988 ◽

Vol 3 (5) ◽

pp. 206-211

Author(s):

PG FitzGerald

Keyword(s):

Gap Junctions ◽

Gap Junction ◽

Molecular Analysis ◽

Tissue Specificity ◽

Structure And Function ◽

Junctional Proteins ◽

And Function ◽

Junction Structure

Gap junctions are nearly ubiquitous structures that ionically and metabolically couple adjacent cells. Molecular analysis of junctional proteins is establishing the presence of families of unique but homologous junctional proteins, opening the door to an explanation of tissue specificity in gap junction structure and function.

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CURRENT VIEWS ON HISTOPHYSIOLOGY OF BRONCHIOLAR EXOCRINE CELLS

Морфология ◽

10.34922/ae.2020.157.1.014 ◽

2020 ◽

pp. 93-97

Author(s):

В. Л. Горячкина ◽

Д. А. Цомартова ◽

Е. В. Черешнева ◽

М. Ю. Иванова ◽

С. Л. Кузнецов

Keyword(s):

Lung Cancer ◽

Lung Tumor ◽

Structure And Function ◽

Clara Cells ◽

Protective Function ◽

Exocrine Cells ◽

Specific Proteins ◽

Harmful Substances ◽

And Function ◽

Metabolism Of Xenobiotics

В обзоре приводятся новые данные о структуре и функции бронхиолярных экзокриноцитов. Впервые нереснитчатые клетки в бронхиолах были описаны ещё А. фон Кёлликером в 1881 г. Детальное изучение этих клеток в бронхиолах человека и кроликов было проведено М. Клара в 1937 г., в честь которого они были названы. В обзоре обсуждаются следующие функции клеток Клара (КК), или бронхиолярных экзокриноцитов: защитная функция, обусловленная секрецией специфических белков, а также жидкого субстрата, располагающегося на поверхности слизистой оболочки; участие в восстановлении повреждённых реснитчатых клеток в качестве своеобразных стволовых (прогениторных) клеток; функция детоксикации вредных веществ, попадающих в лёгкие, а именно: метаболизация ксенобиотиков и канцерогенных веществ; участие в развитии многих форм рака лёгких, источником формирования которых являются бронхиолярные экзокриноциты, включая аденокарциному - наиболее часто диагностируемую опухоль лёгкого. This review provides new data on the structure and function of bronchiolar exocrine cells. The nonciliary cells in the bronchioles were first described by Kolliker as early as in 1881. The detailed study of these cells in human and rabbit bronchioles was carried out by M. Clara in 1973, and the cells were named after him. The review discusses the following functions of Clara cells or bronchiolar exocrine cells: a protective function due to the secretion of specific proteins, as well as a liquid substrate located on the surface of the mucous membrane; participation in the restoration of damaged ciliary cells as a kind of stem (progenitor) cells; the function of detoxification of harmful substances that enter the lungs, namely: the metabolism of xenobiotics and carcinogens; participation in the development of many forms of lung cancer, the source of the formation of which are bronchiolar exocrine cells, including adenocarcinoma, the most commonly diagnosed lung tumor.

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Molecular Analysis of the Structure and Function of the Angiotensin II Type 1 Receptor

Hypertension Research ◽

10.1291/hypres.26.937 ◽

2003 ◽

Vol 26 (12) ◽

pp. 937-943 ◽

Cited By ~ 80

Author(s):

Shin-ichiro MIURA ◽

Keijiro SAKU ◽

Sadashiva S KARNIK

Keyword(s):

Angiotensin Ii ◽

Molecular Analysis ◽

Structure And Function ◽

And Function

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Selective knockdown of hexokinase 2 in rods leads to age-related photoreceptor degeneration and retinal metabolic remodeling

Cell Death and Disease ◽

10.1038/s41419-020-03103-7 ◽

2020 ◽

Vol 11 (10) ◽

Author(s):

Rui Zhang ◽

Weiyong Shen ◽

Jianhai Du ◽

Mark C. Gillies

Keyword(s):

Mass Spectrometry ◽

Aerobic Glycolysis ◽

Tca Cycle ◽

Structure And Function ◽

Photoreceptor Degeneration ◽

Age Related ◽

The Tca Cycle ◽

Metabolic Remodeling ◽

Specific Proteins ◽

And Function

Abstract Photoreceptors, the primary site of phototransduction in the retina, require energy and metabolites to constantly renew their outer segments. They preferentially consume most glucose through aerobic glycolysis despite possessing abundant mitochondria and enzymes for oxidative phosphorylation (OXPHOS). Exactly how photoreceptors balance aerobic glycolysis and mitochondrial OXPHOS to regulate their survival is still unclear. We crossed rhodopsin-Cre mice with hexokinase 2 (HK2)-floxed mice to study the effect of knocking down HK2, the first rate-limiting enzyme in glycolysis, on retinal health and metabolic remodeling. Immunohistochemistry and Western blots were performed to study changes in photoreceptor-specific proteins and key enzymes in glycolysis and the tricarboxylic acid (TCA) cycle. Changes in retinal structure and function were studied by optical coherence tomography and electroretinography. Mass spectrometry was performed to profile changes in 13C-glucose-derived metabolites in glycolysis and the TCA cycle. We found that knocking down HK2 in rods led to age-related photoreceptor degeneration, evidenced by reduced expression of photoreceptor-specific proteins, age-related reductions of the outer nuclear layer, photoreceptor inner and outer segments and impaired electroretinographic responses. Loss of HK2 in rods led to upregulation of HK1, phosphorylation of pyruvate kinase muscle isozyme 2, mitochondrial stress proteins and enzymes in the TCA cycle. Mass spectrometry found that the deletion of HK2 in rods resulted in accumulation of 13C-glucose along with decreased pyruvate and increased metabolites in the TCA cycle. Our data suggest that HK2-mediated aerobic glycolysis is indispensable for the maintenance of photoreceptor structure and function and that long-term inhibition of glycolysis leads to photoreceptor degeneration.

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