From tissue phenotype to proteotype: sensitive protein identification in microdissected tumor tissue.
Correlation of disease phenotype with protein profile (proteotype) is a significant challenge for biomedical research. The main obstacles have been the need to insure sufficient quantities of pure protein sample, the reproducibility of protein display, and rapid and accurate protein identification. We present a modified approach that combines enhanced detection sensitivity with tissue microdissection from frozen primary renal cancer tissues of different histological subtypes, followed by 2D gel analysis and protein identification with MALDI mass spectrometry. We obtained reliable and highly consistent results in phenotypically similar tumors of each individual subtype by performing strict morphological control of the analyzed tumor cells without physical or chemical alteration of the frozen tissue samples. By application of non-oxidizing silver staining, proteins were resolved and identified with high levels of specificity and sensitivity. This new combination of techniques allows not only for sensitive identification of specific protein patterns that correspond to a histological tumor phenotype, but also for identification of specific disease-associated protein targets.
Zhuang
Zhengping
Z
Surgical Neurology Branch, NINDS/NIH, Bethesda, MD 20892, USA.
Huang
Steve
S
Kowalak
Jeff A
JA
Shi
Ying
Y
Lei
Jingqi
J
Furuta
Makoto
M
Lee
Youn-Soo
YS
Lubensky
Irina A
IA
Rodgers
Griffin P
GP
Cornelius
Albert S
AS
Weil
Robert J
RJ
Teh
Bin T
BT
Vortmeyer
Alexander O
AO
D018249:Adenoma, Oxyphilic / Q000235:genetics; D002291:Carcinoma, Papillary / Q000235:genetics; D002292:Carcinoma, Renal Cell / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics; D042282:Microdissection; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology*; D010641:Phenotype; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D009396:Wilms Tumor / Q000235:genetics
D011506:Proteins