• Something wrong with this record ?

Combining chymotrypsin/trypsin digestion to identify hydrophobic proteins from oil bodies

M. Vermachova, Z. Purkrtova, J. Santrucek, P. Jolivet, T. Chardot, M. Kodicek,

. 2014 ; 1072 (-) : 185-98.

Language English Country United States

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc14063883

Oil bodies, lipid-storage organelles, are stabilized by a number of specific proteins. These proteins are very hydrophobic, which complicates their identification by "classical" proteomic protocols using trypsin digestion. Due to the lack of trypsin cleavage sites, the achievable protein coverage is limited or even insufficient for reliable protein identification. To identify such proteins and to enhance their coverage, we introduced a modified method comprising standard three-step procedure (SDS-PAGE, in-gel digestion, and LC-MS/MS analysis). In this method, chymotrypsin, single or in combination with trypsin, was used, which enabled to obtain proteolytic peptides from the hydrophobic regions and to identify new oil bodies' proteins. Our method can be easily applied to identification of other hydrophobic proteins.

References provided by Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc14063883
003      
CZ-PrNML
005      
20140707113447.0
007      
ta
008      
140704s2014 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1007/978-1-62703-631-3_14 $2 doi
035    __
$a (PubMed)24136523
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Vermachova, Martina $u Department of Biochemistry and Microbiology, Institute of Chemical Technology Prague, Prague, Czech Republic.
245    10
$a Combining chymotrypsin/trypsin digestion to identify hydrophobic proteins from oil bodies / $c M. Vermachova, Z. Purkrtova, J. Santrucek, P. Jolivet, T. Chardot, M. Kodicek,
520    9_
$a Oil bodies, lipid-storage organelles, are stabilized by a number of specific proteins. These proteins are very hydrophobic, which complicates their identification by "classical" proteomic protocols using trypsin digestion. Due to the lack of trypsin cleavage sites, the achievable protein coverage is limited or even insufficient for reliable protein identification. To identify such proteins and to enhance their coverage, we introduced a modified method comprising standard three-step procedure (SDS-PAGE, in-gel digestion, and LC-MS/MS analysis). In this method, chymotrypsin, single or in combination with trypsin, was used, which enabled to obtain proteolytic peptides from the hydrophobic regions and to identify new oil bodies' proteins. Our method can be easily applied to identification of other hydrophobic proteins.
650    _2
$a Arabidopsis $x metabolismus $7 D017360
650    _2
$a proteiny huseníčku $x metabolismus $7 D029681
650    _2
$a chromatografie kapalinová $7 D002853
650    _2
$a chymotrypsin $x metabolismus $7 D002918
650    _2
$a elektroforéza v polyakrylamidovém gelu $7 D004591
650    12
$a hydrofobní a hydrofilní interakce $7 D057927
650    12
$a lipidy $7 D008055
650    _2
$a hmotnostní spektrometrie $7 D013058
650    _2
$a organely $x metabolismus $7 D015388
650    _2
$a proteomika $x metody $7 D040901
650    _2
$a trypsin $x metabolismus $7 D014357
655    _2
$a časopisecké články $7 D016428
700    1_
$a Purkrtova, Zita
700    1_
$a Santrucek, Jiri
700    1_
$a Jolivet, Pascale
700    1_
$a Chardot, Thierry
700    1_
$a Kodicek, Milan
773    0_
$w MED00180389 $t Methods in molecular biology (Clifton, N.J.) $x 1940-6029 $g Roč. 1072, č. - (2014), s. 185-98
856    41
$u https://pubmed.ncbi.nlm.nih.gov/24136523 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20140704 $b ABA008
991    __
$a 20140707113735 $b ABA008
999    __
$a ok $b bmc $g 1031367 $s 862615
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2014 $b 1072 $c - $d 185-98 $i 1940-6029 $m Methods in molecular biology $n Methods Mol Biol $x MED00180389
LZP    __
$a Pubmed-20140704
Back

Find record

Citation metrics

Archiving options