GPI Membrane Anchors – The Much Needed link

Proteomic Approaches for GPI-Anchored Protein Analysis

Author(s): Miren J. Omaetxebarria and Felix Elortza

Pp: 53-63 (11)

DOI: 10.2174/978160805123611001010053

* (Excluding Mailing and Handling)

Abstract

GPI-anchored membrane proteins (GPI-APs) are a functionally and structurally diverse protein family present in eukaryote cells. Their common feature is the anchoring mechanism via glycosylphosphatidylinositol (GPI) to a lipid located in the extracellular leaflet of plasma membrane. Following a modification specific proteomic strategy, GPI-APs can be selectively isolated by a combined treatment of membrane fractions with Triton X-114 detergent and phosphatidylinositol-specific phospholipase C and/or D. The released proteins can be further identified by liquid chromatography on-line coupled to a tandem mass spectrometer. Although the method is proven to be highly specific, some non-GPI-proteins are also known to be released. Bioinformatics has been regularly used to discriminate the GPI-APs among all the identified proteins with a high degree of accuracy. Structural characterization of the GPI-anchors has been historically a laborious analytical task. Recently, HILIC (Hydrophilic Interaction Liquid Chromatography) and titanium dioxide enrichment of GPI-modified peptides in combination with mass spectrometry have been used to isolate and analyse GPI-anchors as well as for the determination of the anchor attachment site. The amino acid where the previously synthesized GPI-anchor is attached is named the omega site (ω-site). Moreover, GPI-specific diagnostic ions detected in tandem mass spectra can potentially be used in large-scale proteomic experiments to track GPI-specific peptides in complex mixtures. All the information obtained by the mentioned strategies has been used for the development of an integrated computational and experimental proteomic approach designed for identification of GPI-anchored peptides in MS/MS spectra as well as for ω-site determination.

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      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Trial Access" as access_base_text, CASE j.type WHEN 111 THEN true ELSE false END as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_trail` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 316323591 and j.to_ip >=316323591 )) and ((j.from_date <= 1739847310 and j.to_date >= 1739847310 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 1858) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Subscribed" as access_base_text, false as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_corporate` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 316323591 and j.to_ip >=316323591 )) and ((j.from_date <= 1739847310 and j.to_date >= 1739847310 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 1858) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Token Based Access" as access_base_text, false as terms_popup, "R" as content_download_type, j.uc_id as restricted_user_access_key, "v" as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_token` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 316323591 and j.to_ip >=316323591 )) and ((j.from_date <= 1739847310 and j.to_date >= 1739847310 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 1858)
      580μsalphaeurekaselec_live_10_06_2022UserAccess.php#1266
      Backtrace
      • 13. app/Models/UserAccess/UserAccess.php:1266
      • 14. app/Http/Controllers/ChapterController.php:1549
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select b.banner_link, b.banner_name, b.banner_script, b.banner_remarks, b.banner_file_name, b.banner_placement, bd.content_type, bd.content_id from `tbl_banner` as `b` inner join `tbl_banner_detail` as `bd` on `bd`.`banner_id` = `b`.`banner_id` inner join `users` as `u` on `u`.`id` = `b`.`updated_by` where `b`.`banner_ebook` = 1 and `bd`.`content_type` = 'B' and `bd`.`content_id` = 922 and `b`.`banner_from_date` <= 1739847310 and `b`.`banner_to_date` >= 1739847310 and `b`.`banner_status` = 'A' order by `b`.`banner_id` desc
      1.03msalphaeurekaselec_live_10_06_2022Banner.php#95
      Bindings
      • 0: 1
      • 1: B
      • 2: 922
      • 3: 1739847310
      • 4: 1739847310
      • 5: A
      Backtrace
      • 13. app/Models/Banner.php:95
      • 14. app/Http/helpers.php:404
      • 17. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 18. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    • select count(*) as aggregate from `uc_cart_products` where cart_id='99a79b226a18b83ecf0b44f6d2f1366b'
      210μsalphaeurekaselec_live_10_06_2022Cart.php#271
      Backtrace
      • 15. app/Models/Cart.php:271
      • 16. view::layouts._header:260
      • 18. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 20. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    App\Models\Chapter
    1Chapter.php#?
    App\Models\Keywords
    1Keywords.php#?
    App\Models\Ebook
    1Ebook.php#?
        _token
        7aLEGuGAATIoXSIRYPRddMY94hoRoD6gWSXQYVBx
        _previous
        array:1 [ "url" => "https://alpha.eurekaselect.com/chapter/1858" ]
        _flash
        array:2 [ "old" => [] "new" => [] ]
        uc_cart_id
        99a79b226a18b83ecf0b44f6d2f1366b
        PHPDEBUGBAR_STACK_DATA
        []
        path_info
        /chapter/1858
        status_code
        200
        
        status_text
        OK
        format
        html
        content_type
        text/html; charset=UTF-8
        request_query
        []
        
        request_request
        []
        
        request_headers
        0 of 0
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        request_cookies
        0 of 0
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        response_headers
        0 of 0
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        session_attributes
        0 of 0
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        ClearShow all
        Date ↕MethodURLData
        #12025-02-18 02:55:10GET/chapter/1858112193