scholarly journals Dowling, P.; et al. Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics. Proteomes 2019, 7, 25

Proteomes ◽  
2019 ◽  
Vol 7 (3) ◽  
pp. 28
Author(s):  
Paul Dowling ◽  
Margit Zweyer ◽  
Dieter Swandulla ◽  
Kay Ohlendieck

The authors wish to make the following correction to their paper [...]

Proteomes ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 25 ◽  
Author(s):  
Paul Dowling ◽  
Margit Zweyer ◽  
Dieter Swandulla ◽  
Kay Ohlendieck

The mass spectrometric analysis of skeletal muscle proteins has used both peptide-centric and protein-focused approaches. The term ‘top-down proteomics’ is often used in relation to studying purified proteoforms and their post-translational modifications. Two-dimensional gel electrophoresis, in combination with peptide generation for the identification and characterization of intact proteoforms being present in two-dimensional spots, plays a critical role in specific applications of top-down proteomics. A decisive bioanalytical advantage of gel-based and top-down approaches is the initial bioanalytical focus on intact proteins, which usually enables the swift identification and detailed characterisation of specific proteoforms. In this review, we describe the usage of two-dimensional gel electrophoretic top-down proteomics and related approaches for the systematic analysis of key components of the contractile apparatus, with a special focus on myosin heavy and light chains and their associated regulatory proteins. The detailed biochemical analysis of proteins belonging to the thick and thin skeletal muscle filaments has decisively improved our biochemical understanding of structure-function relationships within the contractile apparatus. Gel-based and top-down proteomics has clearly established a variety of slow and fast isoforms of myosin, troponin and tropomyosin as excellent markers of fibre type specification and dynamic muscle transition processes.


Author(s):  
Dirk Doyle ◽  
Lawrence Benedict ◽  
Fritz Christian Awitan

Abstract Novel techniques to expose substrate-level defects are presented in this paper. New techniques such as inter-layer dielectric (ILD) thinning, high keV imaging, and XeF2 poly etch overflow are introduced. We describe these techniques as applied to two different defects types at FEOL. In the first case, by using ILD thinning and high keV imaging, coupled with focused ion beam (FIB) cross section and scanning transmission electron microscopy (STEM,) we were able to judge where to sample for TEM from a top down perspective while simultaneously providing the top down images giving both perspectives on the same sample. In the second case we show retention of the poly Si short after removal of CoSi2 formation on poly. Removal of the CoSi2 exposes the poly Si such that we can utilize XeF2 to remove poly without damaging gate oxide to reveal pinhole defects in the gate oxide. Overall, using these techniques have led to 1) increased chances of successfully finding the defects, 2) better characterization of the defects by having a planar view perspective and 3) reduced time in localizing defects compared to performing cross section alone.


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