Global analysis of protein activities using proteome chips.
To facilitate studies of the yeast proteome, we cloned 5800 open reading frames and overexpressed and purified their corresponding proteins. The proteins were printed onto slides at high spatial density to form a yeast proteome microarray and screened for their ability to interact with proteins and phospholipids. We identified many new calmodulin- and phospholipid-interacting proteins; a common potential binding motif was identified for many of the calmodulin-binding proteins. Thus, microarrays of an entire eukaryotic proteome can be prepared and screened for diverse biochemical activities. The microarrays can also be used to screen protein-drug interactions and to detect posttranslational modifications.
Zhu
H
H
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Bilgin
M
M
Bangham
R
R
Hall
D
D
Casamayor
A
A
Bertone
P
P
Lan
N
N
Jansen
R
R
Bidlingmaier
S
S
Houfek
T
T
Mitchell
T
T
Miller
P
P
Dean
R A
RA
Gerstein
M
M
Snyder
M
M
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D019151:Peptide Library; D010713:Phosphatidylcholines / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D020543:Proteome; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D019809:Streptavidin / Q000378:metabolism
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D005656:Fungal Proteins; D008081:Liposomes; D008565:Membrane Proteins; D019151:Peptide Library; D010713:Phosphatidylcholines; D010716:Phosphatidylinositols; D010743:Phospholipids; D020543:Proteome; D011993:Recombinant Fusion Proteins; D019809:Streptavidin; D005947:Glucose