PMID- 22486727
OWN - NLM
STAT- MEDLINE
DCOM- 20120910
LR  - 20161125
IS  - 1535-3907 (Electronic)
IS  - 1535-3893 (Linking)
VI  - 11
IP  - 5
DP  - 2012 May 4
TI  - Low temperature stress modulated secretome analysis and purification of 
      antifreeze protein from Hippophae rhamnoides, a Himalayan wonder plant.
PG  - 2684-96
LID - 10.1021/pr200944z [doi]
AB  - Plants' distribution and productivity are adversely affected by low temperature 
      (LT) stress. LT induced proteins were analyzed by 2-DE-nano-LC-MS/MS in shoot 
      secretome of Hippophae rhamnoides (seabuckthorn), a Himalayan wonder shrub. 
      Seedlings were subjected to direct freezing stress (-5  degrees C), cold acclimation 
      (CA), and subzero acclimation (SZA), and extracellular proteins (ECPs) were 
      isolated using vacuum infiltration. Approximately 245 spots were reproducibly 
      detected in 2-DE gels of LT treated secretome, out of which 61 were LT 
      responsive. Functional categorization of 34 upregulated proteins showed 47% 
      signaling, redox regulated, and defense associated proteins. LT induced secretome 
      contained thaumatin like protein and Chitinase as putative antifreeze proteins 
      (AFPs). Phase contrast microscopy with a nanoliter osmometer showed hexagonal ice 
      crystals with 0.13  degrees C thermal hysteresis (TH), and splat assay showed 1.5-fold 
      ice recrystallization inhibition (IRI), confirming antifreeze activity in LT 
      induced secretome. A 41 kDa polygalacturonase inhibitor protein (PGIP), purified 
      by ice adsorption chromatography (IAC), showed hexagonal ice crystals, a TH of 
      0.19  degrees C, and 9-fold IRI activity. Deglycosylated PGIP retained its AFP activity, 
      suggesting that glycosylation is not required for AFP activity. This is the first 
      report of LT modulated secretome analysis and purification of AFPs from 
      seabuckthorn. Overall, these findings provide an insight in probable LT induced 
      signaling in the secretome.
FAU - Gupta, Ravi
AU  - Gupta R
AD  - Molecular Plant Physiology and Proteomics Laboratory, Department of Botany, 
      University of Delhi, Delhi-110007, India.
FAU - Deswal, Renu
AU  - Deswal R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120423
PL  - United States
TA  - J Proteome Res
JT  - Journal of proteome research
JID - 101128775
RN  - 0 (Antifreeze Proteins)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Plant Proteins)
RN  - 53850-34-3 (thaumatin protein, plant)
RN  - EC 1.15.1.1 (Superoxide Dismutase)
RN  - EC 3.2.1.14 (Chitinases)
RN  - EC 4.4.1.5 (Lactoylglutathione Lyase)
SB  - IM
MH  - Acclimatization
MH  - Antifreeze Proteins/chemistry/*isolation & purification
MH  - Chitinases/chemistry/*isolation & purification
MH  - *Cold Temperature
MH  - Electrophoresis, Gel, Two-Dimensional
MH  - Enzyme Activation
MH  - Extracellular Matrix Proteins/chemistry/isolation & purification
MH  - Hippophae/*chemistry/enzymology
MH  - Lactoylglutathione Lyase/chemistry/isolation & purification
MH  - Microscopy, Phase-Contrast
MH  - Osmometry
MH  - Oxidation-Reduction
MH  - Plant Proteins/chemistry/isolation & purification
MH  - Secretory Pathway
MH  - Signal Transduction
MH  - *Stress, Physiological
MH  - Superoxide Dismutase/chemistry/isolation & purification
EDAT- 2012/04/11 06:00
MHDA- 2012/09/11 06:00
CRDT- 2012/04/11 06:00
PHST- 2012/04/11 06:00 [entrez]
PHST- 2012/04/11 06:00 [pubmed]
PHST- 2012/09/11 06:00 [medline]
AID - 10.1021/pr200944z [doi]
PST - ppublish
SO  - J Proteome Res. 2012 May 4;11(5):2684-96. doi: 10.1021/pr200944z. Epub 2012 Apr 
      23.