Cracks in the foundation: keratin filaments and genetic disease

In the past three years, defects in the genes that encode intermediate filament (IF) proteins have been found to be responsible for some inherited skin diseases, and others have been implicated in certain motor neuron diseases and cardiomyopathies. This article reviews how knowledge of IF structure led to the discovery of genetic disorders of IFs, and how the clinical manifestations of these diseases have confirmed the notion that IFs provide the mechanical strength of cells.     

封闭蛋白的结构、功能及其与人类疾病

紧密连接（tight junction,TJ）是脊椎动物细胞间连接的一种主要形式,对介导上皮细胞间的黏合、维持上皮细胞的功能具有重要作用。TJ是由一系列跨膜蛋白和外周蛋白相互作用而形成的一个复杂的蛋白质体系,封闭蛋白（occludin）是构成TJ的主要成分之一。目前,已发现封闭蛋白与许多人类疾病有关。本文仅就封闭蛋白的结构、功能及其与人类疾病的关系做一综述。     

The mTOR pathway and its role in human genetic diseases

The signalling components upstream and downstream of the protein kinase mammalian target of rapamycin (mTOR) are frequently altered in a wide variety of human diseases. Upstream of mTOR key signalling molecules are the small GTPase Ras, the lipid kinase PI3K, the Akt kinase, and the GTPase Rheb, which are known to be deregulated in many human cancers. Mutations in the mTOR pathway component genes TSC1, TSC2, LKB1, PTEN, VHL, NF1 and PKD1 trigger the development of the syndromes tuberous sclerosis, Peutz-Jeghers syndrome, Cowden syndrome, Bannayan-Riley-Ruvalcaba syndrome, Lhermitte-Duclos disease, Proteus syndrome, von Hippel-Lindau disease, Neurofibromatosis type 1, and Polycystic kidney disease, respectively. In addition, the tuberous sclerosis proteins have been implicated in the development of several sporadic tumors and in the control of the cyclin-dependent kinase inhibitor p27, known to be of relevance for several cancers. Recently, it has been recognized that mTOR is regulated by TNF-alpha and Wnt, both of which have been shown to play critical roles in the development of many human neoplasias. In addition to all these human diseases, the role of mTOR in Alzheimer's disease, cardiac hypertrophy, obesity and type 2 diabetes is discussed.     

Biophysical Reviews— 2021, the year that was

The current issue (volume 13 issue 6, 2021) is a Special Issue jointly dedicated to scientific content presented at the 20th triennial IUPAB Congress that was held in conjunction with both the 45th Annual Meeting of the Brazilian Biophysical Society (Sociedade Brasileira de Biofísica - SBBf) and the 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology (Sociedade Brasileira de Bioquímica e Biologia Molecular – SBBq). In addition to describing the scientific and nonscientific content arising from the meeting this sub-editorial also provides a look back at some of the high points for Biophysical Reviews in the year 2021 before going on to describe a number of matters of interest to readers of the journal in relation to the coming year of 2022.

This Editorial marks the last issue for the journal to be published in 2021 – a year that has been characterized by a mixture of hardship, frustration, and of late, (possibly) a slowly developing cautious optimism in relation to the COVID-19 pandemic. Over the last 2 years, the journal has had to rapidly adapt to suddenly altered plans of contributors, as the publication of scientific reviews and organization of conference-based special issues has necessarily taken a back seat to the realities of altered work practices and, in some cases, changed life and career plans. One such major change was directly concerned with the subject of this special issue (SI) on the scientific content associated with the 20th Congress of the IUPAB (International Union for Pure and Applied Biophysics) conducted in concert with the 45th Annual Meeting of the Brazilian Biophysical Society (SBBf) and the 50th Annual Meeting of the Brazilian Biochemical and Molecular Biology Society (SBBq) (Itri et al. 2021). After discussing a few notable features of the SI, this editorial will introduce important developments occurring with the journal that relate to new feature commentaries and Institutional access arrangements. This Editorial will then close with a look back at some of the standout articles of 2021.     

Exploring the role of galectin 3 in kidney function: a genetic approach

Galectin 3 belongs to a family of glycoconjugate-binding proteinsthat participate in cellular homeostasis by modulating cellgrowth, adhesion, and signaling. We studied adult galectin 3null mutant (Gal 3–/–) and wild-type (WT) mice togain insights into the role of galectin 3 in the kidney. Byimmunofluorescence, galectin 3 was found in collecting duct(CD) principal and intercalated cells in some regions of thekidney, as well as in the thick ascending limbs at lower levels.Compared to WT mice, Gal 3–/– mice had ~11% fewerglomeruli (p < 0.04), associated with kidney hypertrophy(p < 0.006). In clearance experiments, urinary chloride excretionwas found to be higher in Gal 3–/– than in WT mice(p < 0.04), but there was no difference in urinary bicarbonateexcretion, in glomerular filtration, or urinary flow rates.Under chronic low sodium diet, Gal 3–/– mice hadlower extracellular fluid (ECF) volume than WT mice (p <0.05). Plasma aldosterone concentration was higher in Gal 3–/–than in WT mice (p < 0.04), which probably caused the observedincrease in -epithelial sodium channel (-ENaC) protein abundancein the mutant mice (p < 0.001). Chronic high sodium dietresulted paradoxically in lower blood pressure (p < 0.01)in Gal 3–/– than in WT. We conclude that Gal 3–/–mice have mild renal chloride loss, which causes chronic ECFvolume contraction and reduced blood pressure levels.     

The role of keratin subfamilies and keratin pairs in the formation of human epidermal intermediate filaments

The four major keratins of normal human epidermis (molecular mass 50, 56.5, 58, and 65-67 kD) can be subdivided on the basis of charge into two subfamilies (acidic 50-kD and 56.5-kD keratins vs. relatively basic 58-kD and 65-67-kD keratins) or subdivided on the basis of co-expression into two "pairs" (50-kD/58-kD keratin pair synthesized by basal cells vs. 56.5-kD/65-67-kD keratin pair expressed in suprabasal cells). Acidic and basic subfamilies were separated by ion exchange chromatography in 8.5 M urea and tested for their ability to reassemble into 10-nm filaments in vitro. The two keratins in either subfamily did not reassemble into 10-nm filaments unless combined with members of the other subfamily. While electron microscopy of acidic and basic keratins equilibrated in 4.5 M urea showed that keratins within each subfamily formed distinct oligomeric structures, possibly representing precursors in filament assembly, chemical cross-linking followed by gel analysis revealed dimers and larger oligomers only when subfamilies were combined. In addition, among the four major keratins, the acidic 50-kD and basic 58-kD keratins showed preferential association even in 8.5 M urea, enabling us to isolate a 50-kD/58-kD keratin complex by gel filtration. This isolated 50-kD/58-kD keratin pair readily formed 10-nm filaments in vitro. These results demonstrate that in tissues containing multiple keratins, two keratins are sufficient for filament assembly, but one keratin from each subfamily is required. More importantly, these data provide the first evidence for the structural significance of specific co-expressed acidic/basic keratin pairs in the formation of epithelial 10-nm filaments.     

Exonic splicing enhancers: mechanism of action, diversity and role in human genetic diseases

Exonic splicing enhancers (ESEs) are discrete sequences within exons that promote both constitutive and regulated splicing. The precise mechanism by which ESEs facilitate the assembly of splicing complexes has been controversial. However, recent studies have provided insights into this question and have led to a new model for ESE function. Other recent work has suggested that ESEs are comprised of diverse sequences and occur frequently within exons. Ominously, these latter studies predict that many human genetic diseases linked to mutations within exons might be caused by the inactivation of ESEs.     

Emerging role of autophagy in kidney function,diseases and aging

Autophagy is a highly conserved process that degrades cellular long-lived proteins and organelles. Accumulating evidence indicates that autophagy plays a critical role in kidney maintenance, diseases and aging. Ischemic, toxic, immunological, and oxidative insults can cause an induction of autophagy in renal epithelial cells modifying the course of various kidney diseases. This review summarizes recent insights on the role of autophagy in kidney physiology and diseases alluding to possible novel intervention strategies for treating specific kidney disorders by modifying autophagy.     

Emerging role of autophagy in kidney function,diseases and aging

Autophagy is a highly conserved process that degrades cellular long-lived proteins and organelles. Accumulating evidence indicates that autophagy plays a critical role in kidney maintenance, diseases and aging. Ischemic, toxic, immunological, and oxidative insults can cause an induction of autophagy in renal epithelial cells modifying the course of various kidney diseases. This review summarizes recent insights on the role of autophagy in kidney physiology and diseases alluding to possible novel intervention strategies for treating specific kidney disorders by modifying autophagy.     

Lipid rafts: structure, function and role in HIV, Alzheimer's and prion diseases

The fluid mosaic model of the plasma membrane has evolved considerably since its original formulation 30 years ago. Membrane lipids do not form a homogeneous phase consisting of glycerophospholipids (GPLs) and cholesterol, but a mosaic of domains with unique biochemical compositions. Among these domains, those containing sphingolipids and cholesterol, referred to as membrane or lipid rafts, have received much attention in the past few years. Lipid rafts have unique physicochemical properties that direct their organisation into liquid-ordered phases floating in a liquid-crystalline ocean of GPLs. These domains are resistant to detergent solubilisation at 4 degrees C and are destabilised by cholesterol- and sphingolipid-depleting agents. Lipid rafts have been morphologically characterised as small membrane patches that are tens of nanometres in diameter. Cellular and/or exogenous proteins that interact with lipid rafts can use them as transport shuttles on the cell surface. Thus, rafts act as molecular sorting machines capable of co-ordinating the spatiotemporal organisation of signal transduction pathways within selected areas ('signalosomes') of the plasma membrane. In addition, rafts serve as a portal of entry for various pathogens and toxins, such as human immunodeficiency virus 1 (HIV-1). In the case of HIV-1, raft microdomains mediate the lateral assemblies and the conformational changes required for fusion of HIV-1 with the host cell. Lipid rafts are also preferential sites of formation for pathological forms of the prion protein (PrPSc) and of the [beta]-amyloid peptide associated with Alzheimer's disease. The possibility of modulating raft homeostasis, using statins and synthetic sphingolipid analogues, offers new approaches for therapeutic interventions in raft-associated diseases.     

The role of arylamine N-acetyltransferases in chronic degenerative diseases: Their possible function in the immune system

Since their discovery, arylamine N-acetyltransferases 1 and 2 (NAT1 and NAT2, respectively) have been associated with the metabolism of xenobiotics. NAT2 is the main factor in the therapeutic success of tuberculosis treatment due to its role in the biotransformation of isoniazid. However, researchers have started to investigate the possible participation of NAT1 and NAT2 (NATs) in carcinogenesis, although the mechanisms have not been elucidated fully. NATs enzymatic activity is essential in some types of cancer, such as breast cancer and acute lymphoblastic leukemia. Whether NAT1 and/or NAT2 participate in insulin resistance level in diabetes mellitus or in the immune system remains to be explored. Therefore, it is clear that its role in cell physiology has more implications than just metabolizing compounds.     

细胞内一种耗能蛋白质降解途径的发现——2004年诺贝尔化学奖工作介绍

2004年诺贝尔化学奖授予Aaron Ciechanover,Avram Hershko和Irwin Rose三位科学家,以表彰他们在上世纪80年代发现了泛素介导的蛋白质降解过程。文章简单介绍了该现象的科学发现历程,并讨论了该科学发现历程给予我们的启示。     

A role for phosphorylation in the dynamics of keratin intermediate filaments

Keratins undergo highly dynamic events in the epithelial cells that express them. These dynamic changes have been associated with important cell processes. We have studied the possible role of keratin phosphorylation-dephosphorylation processes in the control of these dynamic events. Drugs that affect the protein phosphorylation metabolism (activators or inhibitors of protein kinases or protein phosphatases) have been used in two different dynamic experimental systems. First, the behaviour of keratins after the formation of cell heterokaryons, and second, the assembly of a newly synthesised keratin after transfection into the pre-existing keratin cytoskeleton. The main difference between these two systems stems on the alteration of the amount of keratin polypeptides present in the cells, since in heterokaryons this amount was unaltered whilst in transfection experiments there is an increase due to the presence of the transfected protein. We observed in both systems that the inhibition of protein kinases led to a delayed dynamic behaviour of the keratin polypeptides. On the contrary, the inhibition of protein phosphatases by okadaic acid or the activation of protein kinases by phorbol esters promoted a substantial increase in the kinetics of these processes. Biochemical studies demonstrate that this behavioural changes can be correlated with changes in the phosphorylation state of the keratin polypeptides. As a whole, present results indicate that the highly dynamic properties of the keratin polypeptides can be modulated by phosphorylation.     

p62蛋白的分子功能及其在疾病中的研究进展

螯合体1(SQSTM1/p62)是一种选择性自噬接头蛋白,在清除待降解蛋白、维持细胞内蛋白质稳态中发挥重要的调控作用。p62蛋白具有多个功能结构域,介导与多种蛋白质发生相互作用进而精确调节特定的信号通路,从而将p62蛋白与氧化防御系统、炎症反应和营养感知等重要生命过程联系起来。研究表明p62的突变或者表达异常与多种疾病的发生发展过程密切相关,包括神经退行性疾病、肿瘤、感染性疾病、遗传性疾病以及慢性疾病等。本文综述了p62蛋白的结构特征、分子功能,并系统介绍其在蛋白质稳态和信号通路调控中的多种功能,总结了p62在疾病发生发展中的复杂性与多面性,以期为p62蛋白的功能与相关疾病研究提供参考。