Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers
The outstanding material properties of spider dragline silk fibers have been attributed to two spidroins, major ampullate spidroins 1 and 2 (MaSp1 and MaSp2). Although dragline silk fibers have been treated with different chemical solvents to elucidate the relationship between protein structure and...
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doaj-89ab8c3039134504b84acb95f790d4e52020-11-25T02:32:25ZengMDPI AGInternational Journal of Molecular Sciences1422-00672016-09-01179153710.3390/ijms17091537ijms17091537Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk FibersCamille Larracas0Ryan Hekman1Simmone Dyrness2Alisa Arata3Caroline Williams4Taylor Crawford5Craig A. Vierra6Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USADepartment of Biological Sciences, University of the Pacific, Stockton, CA 95211, USAThe outstanding material properties of spider dragline silk fibers have been attributed to two spidroins, major ampullate spidroins 1 and 2 (MaSp1 and MaSp2). Although dragline silk fibers have been treated with different chemical solvents to elucidate the relationship between protein structure and fiber mechanics, there has not been a comprehensive proteomic analysis of the major ampullate (MA) gland, its spinning dope, and dragline silk using a wide range of chaotropic agents, inorganic salts, and fluorinated alcohols to elucidate their complete molecular constituents. In these studies, we perform in-solution tryptic digestions of solubilized MA glands, spinning dope and dragline silk fibers using five different solvents, followed by nano liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis with an Orbitrap Fusion™ Tribrid™. To improve protein identification, we employed three different tryptic peptide fragmentation modes, which included collision-induced dissociation (CID), electron transfer dissociation (ETD), and high energy collision dissociation (HCD) to discover proteins involved in the silk assembly pathway and silk fiber. In addition to MaSp1 and MaSp2, we confirmed the presence of a third spidroin, aciniform spidroin 1 (AcSp1), widely recognized as the major constituent of wrapping silk, as a product of dragline silk. Our findings also reveal that MA glands, spinning dope, and dragline silk contain at least seven common proteins: three members of the Cysteine-Rich Protein Family (CRP1, CRP2 and CRP4), cysteine-rich secretory protein 3 (CRISP3), fasciclin and two uncharacterized proteins. In summary, this study provides a proteomic blueprint to construct synthetic silk fibers that most closely mimic natural fibers.http://www.mdpi.com/1422-0067/17/9/1537dragline silkmajor ampullateproteomicsblack widow spidercob-weaverspidroin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Camille Larracas Ryan Hekman Simmone Dyrness Alisa Arata Caroline Williams Taylor Crawford Craig A. Vierra |
spellingShingle |
Camille Larracas Ryan Hekman Simmone Dyrness Alisa Arata Caroline Williams Taylor Crawford Craig A. Vierra Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers International Journal of Molecular Sciences dragline silk major ampullate proteomics black widow spider cob-weaver spidroin |
author_facet |
Camille Larracas Ryan Hekman Simmone Dyrness Alisa Arata Caroline Williams Taylor Crawford Craig A. Vierra |
author_sort |
Camille Larracas |
title |
Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers |
title_short |
Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers |
title_full |
Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers |
title_fullStr |
Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers |
title_full_unstemmed |
Comprehensive Proteomic Analysis of Spider Dragline Silk from Black Widows: A Recipe to Build Synthetic Silk Fibers |
title_sort |
comprehensive proteomic analysis of spider dragline silk from black widows: a recipe to build synthetic silk fibers |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2016-09-01 |
description |
The outstanding material properties of spider dragline silk fibers have been attributed to two spidroins, major ampullate spidroins 1 and 2 (MaSp1 and MaSp2). Although dragline silk fibers have been treated with different chemical solvents to elucidate the relationship between protein structure and fiber mechanics, there has not been a comprehensive proteomic analysis of the major ampullate (MA) gland, its spinning dope, and dragline silk using a wide range of chaotropic agents, inorganic salts, and fluorinated alcohols to elucidate their complete molecular constituents. In these studies, we perform in-solution tryptic digestions of solubilized MA glands, spinning dope and dragline silk fibers using five different solvents, followed by nano liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis with an Orbitrap Fusion™ Tribrid™. To improve protein identification, we employed three different tryptic peptide fragmentation modes, which included collision-induced dissociation (CID), electron transfer dissociation (ETD), and high energy collision dissociation (HCD) to discover proteins involved in the silk assembly pathway and silk fiber. In addition to MaSp1 and MaSp2, we confirmed the presence of a third spidroin, aciniform spidroin 1 (AcSp1), widely recognized as the major constituent of wrapping silk, as a product of dragline silk. Our findings also reveal that MA glands, spinning dope, and dragline silk contain at least seven common proteins: three members of the Cysteine-Rich Protein Family (CRP1, CRP2 and CRP4), cysteine-rich secretory protein 3 (CRISP3), fasciclin and two uncharacterized proteins. In summary, this study provides a proteomic blueprint to construct synthetic silk fibers that most closely mimic natural fibers. |
topic |
dragline silk major ampullate proteomics black widow spider cob-weaver spidroin |
url |
http://www.mdpi.com/1422-0067/17/9/1537 |
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