scholarly journals Heterogeneity of proteome dynamics between connective tissue phases of adult tendon

2020 ◽  
Author(s):  
Deborah Simpson ◽  
Howard Choi ◽  
Ding Wang ◽  
Mark Prescott ◽  
Andrew A. Pitsillides ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Extracellular Matrix ◽  
Connective Tissue ◽  
Protein Turnover ◽  
Tissue Homeostasis ◽  
Turnover Rates ◽  
Connective Tissues ◽  
Isotope Labelling ◽  
Tissue Integrity ◽  
Tissue Proteome

AbstractMaintenance of connective tissue integrity is fundamental to sustain function, requiring protein turnover to repair damaged tissue. However, connective tissue proteome dynamics remain largely undefined, as do differences in turnover rates of individual proteins in the collagen and glycoprotein phases of connective tissue extracellular matrix (ECM). Here, we investigate proteome dynamics in the collagen and glycoprotein phases of connective tissues by exploiting the spatially distinct fascicular (collagen-rich) and interfascicular (glycoprotein-rich) ECM phases of tendon. Using isotope labelling, mass spectrometry and bioinformatics, we calculate turnover rates of individual proteins within rat Achilles tendon and its ECM phases. Our results demonstrate complex proteome dynamics in tendon, with ~1000-fold differences in protein turnover rates, and overall faster protein turnover within the glycoprotein-rich interfascicular matrix compared to the collagen-rich fascicular matrix. These data provide insights into the complexity of proteome dynamics in tendon, likely required to maintain tissue homeostasis.

scholarly journals Heterogeneity of proteome dynamics between connective tissue phases of adult tendon

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Howard Choi ◽  
Deborah Simpson ◽  
Ding Wang ◽  
Mark Prescott ◽  
Andrew A Pitsillides ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Extracellular Matrix ◽  
Connective Tissue ◽  
Protein Turnover ◽  
Tissue Homeostasis ◽  
Turnover Rates ◽  
Connective Tissues ◽  
Isotope Labelling ◽  
Tissue Integrity ◽  
Tissue Proteome

Maintenance of connective tissue integrity is fundamental to sustain function, requiring protein turnover to repair damaged tissue. However, connective tissue proteome dynamics remain largely undefined, as do differences in turnover rates of individual proteins in the collagen and glycoprotein phases of connective tissue extracellular matrix (ECM). Here, we investigate proteome dynamics in the collagen and glycoprotein phases of connective tissues by exploiting the spatially distinct fascicular (collagen-rich) and interfascicular (glycoprotein-rich) ECM phases of tendon. Using isotope labelling, mass spectrometry and bioinformatics, we calculate turnover rates of individual proteins within rat Achilles tendon and its ECM phases. Our results demonstrate complex proteome dynamics in tendon, with ~1000 fold differences in protein turnover rates, and overall faster protein turnover within the glycoprotein-rich interfascicular matrix compared to the collagen-rich fascicular matrix. These data provide insights into the complexity of proteome dynamics in tendon, likely required to maintain tissue homeostasis.

scholarly journals Selective association of an endogenous lectin with connective tissues

1985 ◽  
Vol 73 (1) ◽  
pp. 347-359
Author(s):  
J.W. Catt ◽  
F.L. Harrison
Keyword(s):  
Extracellular Matrix ◽  
Connective Tissue ◽  
Heart Tissue ◽  
Hair Follicles ◽  
Connective Tissues ◽  
Alveolar Cells ◽  
Tissue Sections ◽  
Liver And Kidney ◽  
Endogenous Lectins ◽  
Or Organization

Using indirect immunofluorescence we have localized an endogenous beta-galactoside-specific lectin in resin-embedded rabbit tissue sections. The pattern of lectin distribution correlates well with biochemical estimations of lectin levels, being abundant in intestine, lung and heart tissue and relatively less abundant in skeletal muscle, liver and kidney. In all tissues lectin is found in connective tissue associated with fibroblasts and the extracellular matrix, and at the periphery of morphologically recognizable smooth muscle cells. The lectin is abundant in skin, intestine and blood vessels, where connective tissue forms the tissue architecture. It is also abundant in heart, where it is particularly associated with the capillaries and lung, where it is also found in alveolar cells. Discrete localization of lectin occurs in areas of connective tissue where epithelial elements are differentiating, such as the crypts of Lieberkuhns in the small intestine and hair follicles in the skin. From these observations we suggest that in cells of mesenchymal origin these endogenous lectins may play a role in the elaboration or organization of the extracellular matrix that regulates tissue differentiation in a number of embryonic and adult tissues.

scholarly journals EGR1 Transcription Factor is a Multifaceted Regulator of Matrix Production in Tendons and Other Connective Tissues

2020 ◽  
Vol 21 (5) ◽  
pp. 1664 ◽  
Author(s):  
Emmanuelle Havis ◽  
Delphine Duprez
Keyword(s):  
Extracellular Matrix ◽  
Transcription Factor ◽  
Connective Tissue ◽  
Transcriptional Activity ◽  
Tendon Repair ◽  
Regulatory Elements ◽  
Biological Functions ◽  
Connective Tissues ◽  
Post Translational Modifications ◽  
Matrix Production

Although the transcription factor EGR1 is known as NGF1-A, TIS8, Krox24, zif/268, and ZENK, it still has many fewer names than biological functions. A broad range of signals induce Egr1 gene expression via numerous regulatory elements identified in the Egr1 promoter. EGR1 is also the target of multiple post-translational modifications, which modulate EGR1 transcriptional activity. Despite the myriad regulators of Egr1 transcription and translation, and the numerous biological functions identified for EGR1, the literature reveals a recurring theme of EGR1 transcriptional activity in connective tissues, regulating genes related to the extracellular matrix. Egr1 is expressed in different connective tissues, such as tendon (a dense connective tissue), cartilage and bone (supportive connective tissues), and adipose tissue (a loose connective tissue). Egr1 is involved in the development, homeostasis, and healing processes of these tissues, mainly via the regulation of extracellular matrix. In addition, Egr1 is often involved in the abnormal production of extracellular matrix in fibrotic conditions, and Egr1 deletion is seen as a target for therapeutic strategies to fight fibrotic conditions. This generic EGR1 function in matrix regulation has little-explored implications but is potentially important for tendon repair.

scholarly journals Protein turnover measurement using selected reaction monitoring-mass spectrometry (SRM-MS)

2016 ◽  
Vol 374 (2079) ◽  
pp. 20150362 ◽  
Author(s):  
Stephen W. Holman ◽  
Dean E. Hammond ◽  
Deborah M. Simpson ◽  
John Waters ◽  
Jane L. Hurst ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Turnover ◽  
Tricarboxylic Acid Cycle ◽  
High Mass ◽  
Selected Reaction Monitoring ◽  
Reaction Monitoring ◽  
Turnover Rates ◽  
Mass Spectrometers ◽  
High Mass Resolution ◽  
Sensitivity And Selectivity

Protein turnover represents an important mechanism in the functioning of cells, with deregulated synthesis and degradation of proteins implicated in many diseased states. Therefore, proteomics strategies to measure turnover rates with high confidence are of vital importance to understanding many biological processes. In this study, the more widely used approach of non-targeted precursor ion signal intensity (MS1) quantification is compared with selected reaction monitoring (SRM), a data acquisition strategy that records data for specific peptides, to determine if improved quantitative data would be obtained using a targeted quantification approach. Using mouse liver as a model system, turnover measurement of four tricarboxylic acid cycle proteins was performed using both MS1 and SRM quantification strategies. SRM outperformed MS1 in terms of sensitivity and selectivity of measurement, allowing more confident determination of protein turnover rates. SRM data are acquired using cheaper and more widely available tandem quadrupole mass spectrometers, making the approach accessible to a larger number of researchers than MS1 quantification, which is best performed on high mass resolution instruments. SRM acquisition is ideally suited to focused studies where the turnover of tens of proteins is measured, making it applicable in determining the dynamics of proteins complexes and complete metabolic pathways. This article is part of the themed issue ‘Quantitative mass spectrometry’.

scholarly journals Vertebrogenous syndrome, lumbosacral spine against the backdrop of connective tissue dysplasia in athletes

2016 ◽  
pp. 18-24
Author(s):  
A. Pyantkovsky
Keyword(s):  
Extracellular Matrix ◽  
Connective Tissue ◽  
Postnatal Period ◽  
Tissue Homeostasis ◽  
Lumbosacral Spine ◽  
Duration Of Treatment ◽  
Disc Protrusion ◽  
Anatomical Changes ◽  
Connective Tissue Dysplasia ◽  
Pain Analysis

In Ukraine, according to WHO, the disease of the spine is the fourth after cardiovascular system, oncology and pathology of diabetes. Dysplasia of connective tissue (DST) - a violation of the structure of the connective tissue in the embryonic and postnatal period as a result of genetically modified fibrylohenez extracellular matrix, leading to frustration on tissue homeostasis levels prohradiyentnym course. A clinical and neurological examination of 90 patients with an average age of 31,5 ± 13,5 g. оf vertebral syndrome, lumbosacral spine. The results of the clinical study demonstrated that patients with vertebral syndrome, lumbosacral spine against the background displays GOST significantly (p <0,05) more common intervertebral disc protrusion, more pronounced changes in yellow ties, in the form of thickening. Concomitant diseases (manifestations DST) scoliosis, kyphosis, kyphoscoliosis, hiperruhlyvist joints, tendency to dislocation, stretching ligaments joints - significantly (p <0,05) increase the duration of treatment and the severity of pain. Analysis of the survey data and monitoring of patients in the dynamics of the treatment showed that patients with vertebral syndrome, lumbosacral spine against the background displays GOST significantly (p <0,05) more likely to have anatomical changes in the lumbosacral spine, than in patients without evidence of DST.

scholarly journals Long-lived metabolic enzymes in the crystalline lens identified by pulse-labeling of mice and mass spectrometry

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Pan Liu ◽  
Seby Louis Edassery ◽  
Laith Ali ◽  
Benjamin R Thomson ◽  
Jeffrey N Savas ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Turnover ◽  
Crystalline Lens ◽  
Nuclear Proteins ◽  
Turnover Rates ◽  
Fiber Cells ◽  
Oxidation Reduction ◽  
Age Related ◽  
Health And Disease ◽  
Biochemical State

The lenticular fiber cells are comprised of extremely long-lived proteins while still maintaining an active biochemical state. Dysregulation of these activities has been implicated in diseases such as age-related cataracts. However, the lenticular protein dynamics underlying health and disease is unclear. We sought to measure the global protein turnover rates in the eye using nitrogen-15 labeling of mice and mass spectrometry. We measured the 14N/15N-peptide ratios of 248 lens proteins, including Crystallin, Aquaporin, Collagen and enzymes that catalyze glycolysis and oxidation/reduction reactions. Direct comparison of lens cortex versus nucleus revealed little or no 15N-protein contents in most nuclear proteins, while there were a broad range of 14N/15N ratios in cortex proteins. Unexpectedly, like Crystallins, many enzymes with relatively high abundance in nucleus were also exceedingly long-lived. The slow replacement of these enzymes in spite of young age of mice suggests their potential roles in age-related metabolic changes in the lens.

A mass spectrometry workflow for measuring protein turnover rates in vivo

2019 ◽  
Vol 14 (12) ◽  
pp. 3333-3365 ◽  
Author(s):  
Mihai Alevra ◽  
Sunit Mandad ◽  
Till Ischebeck ◽  
Henning Urlaub ◽  
Silvio O. Rizzoli ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Turnover ◽  
Turnover Rates

Vertebrogenous syndrome, lumbosacral spine against the backdrop of connective tissue dysplasia in athletes

2018 ◽  
pp. 27-31
Author(s):  
A. Pyantkovsky
Keyword(s):  
Extracellular Matrix ◽  
Connective Tissue ◽  
Postnatal Period ◽  
Tissue Homeostasis ◽  
Lumbosacral Spine ◽  
Duration Of Treatment ◽  
Disc Protrusion ◽  
Anatomical Changes ◽  
Connective Tissue Dysplasia ◽  
Pain Analysis

In Ukraine, according to WHO, the disease of the spine is the fourth after cardiovascular system, oncology and pathology of diabetes. Dysplasia of connective tissue (DST) - a violation of the structure of the connective tissue in the embryonic and postnatal period as a result of genetically modified fibrylohenez extracellular matrix, leading to frustration on tissue homeostasis levels prohradiyentnym course. A clinical and neurological examination of 90 patients with an average age of 31,5 ± 13,5 g. оf vertebral syndrome, lumbosacral spine. The results of the clinical study demonstrated that patients with vertebral syndrome, lumbosacral spine against the background displays GOST significantly (p <0,05) more common intervertebral disc protrusion, more pronounced changes in yellow ties, in the form of thickening. Concomitant diseases (manifestations DST) scoliosis, kyphosis, kyphoscoliosis, hiperruhlyvist joints, tendency to dislocation, stretching ligaments joints - significantly (p <0,05) increase the duration of treatment and the severity of pain. Analysis of the survey data and monitoring of patients in the dynamics of the treatment showed that patients with vertebral syndrome, lumbosacral spine against the background displays GOST significantly (p <0,05) more likely to have anatomical changes in the lumbosacral spine, than in patients without evidence of DST.

scholarly journals An Approach to Measuring Protein Turnover in Human Induced Pluripotent Stem Cell Organoids by Mass Spectrometry

2021 ◽  
Author(s):  
Anthony Duchesne ◽  
Jing Dong ◽  
Andrew N. Bayne ◽  
Nguyen-Vi Mohamed ◽  
Wei Yi ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Culture ◽  
Protein Turnover ◽  
Isotope Labeling ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Turnover Rates ◽  
Cell Type Specificity ◽  
Induced Pluripotent

AbstractPatient-derived organoids from induced pluripotent stem cells have emerged as a model for studying human diseases beyond conventional two-dimensional (2D) cell culture. Briefly, these three-dimensional organoids are highly complex, capable of self-organizing, recapitulate cellular architecture, and have the potential to model diseases in complex organs, such as the brain. For example, the hallmark of Parkinson’s disease - proteostatic dysfunction leading to the selective death of neurons in the substantia nigra - present a subtle distinction in cell type specificity that is simply lost in 2D cell culture models. As such, the development of robust methods to study global proteostasis and protein turnover in organoids will remain a critical need as organoid models evolve. To solve this problem, we have designed a workflow to extract proteins from organoids and measure global protein turnover using mass spectrometry and stable isotope labeling using amino acids in cell culture (SILAC). This allowed us to measure the turnover rates of 844 proteins and compare protein turnover to previously reported data in primary cell cultures and in vivo models. Taken together, this method will facilitate the study of proteostasis in organoid models of human disease and will provide an analytical and statistical framework to measure protein turnover in organoids of all cell types.

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