Development of peptide aptamer microarrays for detection of HPV16 oncoproteins in cell extracts.
Protein microarrays represent an emerging technology that promises to facilitate high-throughput proteomics. The major goal of this technology is to employ peptides, full-length proteins, antibodies, and small molecules to simultaneously screen thousands of targets for potential protein-protein interactions or modifications of the proteome. This article describes the performance of a set of peptide aptamers specific for the human papillomavirus (HPV) type 16 oncoproteins E6 and E7 in a microarray format. E6 and E7 peptide aptamer microarrays were probed with fluorescence-labeled lysates generated from HPV-infected cervical keratinocytes expressing both E6 and E7 oncoproteins. Peptide aptamer microarrays are shown to detect low levels of E6 and E7 proteins. Peptide aptamers specific for cellular proteins included on these microarrays suggested that expression of CDK2, CDK4, and BCL-6 may be affected by HPV infection and genome integration. We conclude that peptide aptamer microarrays represent a promising tool for proteomics and may be of value in biological and clinical investigations of cervical carcinogenesis.
MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge CB2 0XZ, UK.
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Laurenson
M R
MR
Pett
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Hoppe-Seyler
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Denk
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Hoppe-Seyler
N
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Coleman
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Ko Ferrigno
D052158:Aptamers, Peptide / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry*; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D052162:Human papillomavirus 16 / Q000302:isolation & purification*; D006801:Humans; D015603:Keratinocytes; D015513:Oncogene Proteins / Q000737:chemistry / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000032:analysis*; D050725:Papillomavirus E7 Proteins / Q000737:chemistry / Q000378:metabolism; D030361:Papillomavirus Infections / Q000821:virology; D040081:Protein Array Analysis / Q000379:methods*; D012097:Repressor Proteins / Q000032:analysis*; D002583:Uterine Cervical Neoplasms / Q000821:virology
D052158:Aptamers, Peptide; D002457:Cell Extracts; C068651:E6 protein, Human papillomavirus type 16; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16