Hemolymph proteome at the fifth instar larval stage of Manduca sexta

 

Proteomics is a systematic study of proteins present in a cell, tissue, organ, or organism at a particular moment during the life cycle that began with classical two-dimensional gel electrophoresis (2-DGE). Advancements in 2-DGE during the 1990`s along with a series of tremendous technological developments in mass spectrometry (MS), a core technology, has revolutionized proteomics. Proteomics is exerting its influence on biological function of genes and genomes in the era of functional genomics, and for this reason yeast, bacterial, and mammalian systems are the best examples.

 

Manduca sexta is an excellent insect model for studying insect physiology, including hemolymph proteins. Larvae stages of this insect are highly damaging to tobacco leaves causing drastic decrease in crop yield. Investigation on the larval biology should help in controlling its destructive potential and thus increasing crop yields. The hemolymph is the source of its immunity to disease and environmental factors, and which invariably involves protein components. We can probably discover novel useful bioactive proteins (peptides) from the hemolymph.

 

Here we apply two complementary proteomics approaches, 1- and 2-DGE coupled with Edman sequencing of the N-terminal amino acids and tandem MS to investigate the hemolymph proteome at the fifth instar larval stage of M. sexta. This study reports identification of several novel proteins in hemolymph along with a number of proteins previously reported during the 4 decades of independent and targeted studies. Use of both 1-DGE and 2-DGE based approaches helped in identification of 123 proteins, representing 58 non-redundant proteins. A high-resolution 2-D gel reference map was also established for future comparative proteomics analysis at other stages of the M. sexta life cycle.

 

The result of three experiments is shown:

@ 1) 1-DGE and MS (PDF)

@ 2) 1-DGE and N-terminal sequence (PDF)

@ 3) 2-D map and MS (PDF)

 

The data are described in a paper, gSystematic investigation of hemolymph proteome of Manduca sexta at the fifth instar larvae stage using one- and two-dimensional proteomics platformsh (Furusawa et al., 2007 submitted)