The human protease CLIP-CHIP: Genomic analysis of all 715 human protease and inhibitor gene...
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The human protease CLIP-CHIP: Genomic analysis of all 715 human protease and inhibitor gene transcripts in human breast carcinoma Reinhild Kappelhoff,Tom Ewart+, Xose S. Puente*, Arun Seth+, Carlos López-Otín* and Christopher M. Overall† Departments of Biochemistry and Molecular Biology†, Oral Biological and Medical Sciences, University of British Columbia, Vancouver, BC, Canada ([email protected]); +Department of Molecular Pathology, Sunnybrook & Women's College Health Sciences Centre, Toronto, ON, Canada, *Departamento de Bioquimica y Biologia Molecular, Universidad de Oviedo, Oviedo, Spain Introduction • The availability of the human genome sequence has enabled the identification the degradome—the complete repertoire of proteases produced in man. By using primary information retrieved from public and private sequencing projects, together with data from the MEROPS, InterPro and Ensembl databases we have annotated a total of 558 genes encoding proteases or protease-homologs in the human genome and 157 inhibitors (Xose S. Puente, Luis M. Sanchez, Christopher M. Overall, and Carlos Lopez-Otin, Nature Rev Genetics 4, 544-558, 2003). • To date there are no complete protease microarray chips reported. Hybridization, scanning and analyzing the CLIP-CHIP • Hybridization of labeled cDNA to the microarray was at 25C for 16 h in 5x SSC, 0.3 % SDS. Washing steps were carried out at 42C. • The CLIP-CHIP was scanned in an Affymetrix 428 (MWG) microarray laser scanner using 532 nm for the cyanine 3 channel and 635 nm for the cyanine 5 channel. Photo multiplier gains were set individually for each channel. Conclusions and outlook • The development of the CLIP-CHIP represents the first complete dedicated human protease and inhibitor chip reported to date. • The CLIP-CHIP will prove to be an extremely valuable and versatile tool for the genomic analysis of proteolytic function in vivo. • The advantage of using a custom glass chip is that new protease or inhibitor oligonucleotides can be added in an agile manner at each chip printing for minimal cost. • Moreover, having a dedicated subarray containing oligonucleotides relevant to the experimental question being addressed further focuses the experiment without distraction from analyses of many thousands of genes that is possible in larger arrays or the multiple 20mer oligonucleotides per gene necessary for the Affymetrix chips. • Work is in progress to analyze RNA from an additional 50 breast cancer tissues of different disease stages to analyze their protease and inhibitor expression profiles. The CLIP-CHIP A dedicated human protease chip C L I P C H I P A1 A2 A3 A4 A5 K1 K2 A6 A7 A8 A9 A10 A11 A12 A13 A14 A15 N1 A16 A17 A18 A19 A20 A21 C1 C2 C3 C4 C5 K3 K4 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 K1 K2 C16 C17 C18 C19 C20 C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 C32 C33 C34 C35 C36 C37 K3 K4 C38 C39 C40 C41 C42 C43 C44 C45 C46 C47 C48 C49 C50 C51 C52 C53 N2 N3 C54 C55 C56 C57 C58 C59 C60 C61 C62 C63 C64 C65 C66 C67 C68 C69 C70 C71 C72 C73 C74 C75 N4 N5 C76 C77 C78 C79 C80 C81 C82 C83 C84 C85 C86 C87 C88 C89 C90 C91 C92 C93 C94 C95 C96 C97 C98 C99 C100 C101 C102 C103 C104 C105 C106 C107 C108 C109 C110 C111 C112 C113 C114 C115 K5 K6 C116 C117 C118 C119 C120 C121 C122 C123 C124 C125 K5 K6 C126 C127 C128 C129 C130 N6 C131 C132 C133 C134 K7 K8 C135 C136 C137 C138 C139 C140 C141 C142 C143 C144 K7 K8 C145 C146 C147 C148 N7 M1 M2 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 M13 M14 M15 M16 M17 M18 M19 M20 M21 M22 M23 M24 M25 M26 M27 M28 M29 M30 M31 M32 M33 M34 M35 M36 M37 M38 M39 M40 M41 M42 M43 M44 M45 M46 M47 M48 M49 M50 M51 M52 M53 M54 M55 M56 M57 M58 M59 M60 M61 M62 M63 M64 M65 M66 M67 M68 M69 M70 M71 M72 M73 M74 M75 M76 M77 K12 K9 M78 M79 M80 M81 M82 M83 M84 M85 M86 M87 K12 K9 M88 M89 M90 M91 M92 M93 M94 M95 M96 M97 K10 K11 M98 M99 M100 M101 M102 M103 M104 M105 M106 M107 K10 K11 M108 M109 M110 M111 M112 M113 M114 M115 M116 M117 M118 M119 M120 M121 M122 M123 M124 M125 M126 M127 M128 M129 M130 M131 M132 M133 M134 M135 M136 M137 M138 M139 M140 M141 M142 M143 M144 M145 M146 M147 M148 M149 M150 M151 M152 M153 M154 M155 M156 M157 M158 M159 M160 M161 M162 M163 M164 M165 M166 M167 M168 M169 M170 M171 M172 M173 M174 M175 M176 M177 M178 M179 M180 M181 M182 M183 M184 S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12 S13 S14 S15 S16 S17 S18 S19 S20 S21 S22 S23 S24 S25 S26 S27 S28 S29 S30 S31 S32 S33 S34 S35 S36 S37 S38 S39 S40 S41 S42 S43 S44 S45 S46 S47 S48 S49 S50 S51 S52 S53 S54 S55 S56 S57 S58 S59 S60 S61 S62 S63 S64 S65 S66 S67 S68 S69 S70 S71 S72 S73 S74 S75 S76 S77 K1 K2 S78 S79 S80 S81 S82 S83 S84 S85 S86 S87 K1 K2 S88 S89 S90 S91 S92 S93 S94 S95 S96 S97 K3 K4 S98 S99 S100 S101 S102 S103 S104 S105 S106 S107 K3 K4 S108 S109 S110 S111 S112 S113 S114 S115 S116 S117 S118 S119 S120 S121 S122 S123 S124 S125 S126 S127 S128 S129 S130 S131 S132 S133 S134 S135 S136 S137 S138 S139 S140 S141 S142 S143 S144 S145 S146 S147 S148 S149 S150 S151 S152 S153 S154 S155 S156 S157 S158 S159 S160 S161 S162 S163 S164 S165 S166 S167 S168 S169 S170 S171 S172 S173 S174 S175 S176 S177 K5 K6 K7 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16 T17 T18 T19 T20 T21 T22 T23 T24 T25 T26 T27 T28 I1 I2 I3 I4 I5 I6 I7 I8 I9 I10 I11 I12 I13 I14 I15 I16 I17 I18 I19 I20 I21 I22 I23 I24 I25 I26 I27 I28 I29 I30 I31 I32 I33 I34 I35 I36 I37 I38 I39 I40 I41 I42 I43 I44 I45 I46 I47 I48 I49 I50 I51 I52 I53 K12 K9 I54 I55 I56 I57 I58 I59 I60 I61 I62 I63 K12 K9 I64 I65 I66 I67 I68 I69 I70 I71 I72 I73 K10 K11 I74 I75 I76 I77 I78 I79 I80 I81 I82 I83 K10 K11 I84 I85 I86 I87 I88 I89 I90 I91 I92 I93 I94 I95 I96 I97 I98 I99 I100 I101 I102 I103 I104 I105 I106 I107 I108 I109 I110 I111 I112 I113 I114 I115 I116 I117 I118 I119 I120 I121 I122 I123 I124 I125 I126 I127 I128 I129 I130 I131 I132 I133 I134 I135 I136 I137 I138 I139 I140 I141 I142 I143 I144 I145 I146 I147 I148 I149 I150 I151 I152 I153 I154 I155 I156 I157 K5 K6 K7 Serine (177) Threonine (28) Inhibitor s (157) Aspartic (21) Cysteine (148) Metallo (184) (+) Controls (46) (-) Controls (7) (A) (B) Inhibitors Serine Cysteine Metallo Aspartate Threonine (C) low expression level high Figure 1. The human protease CLIP-CHIP. (A) Overview of the CLIP-CHIP microarray layout. The human microarray is spotted in duplicate (1, 2). In addition 192 control oligonuclotides and a dedicated subarray with 217 breast cancer related and prognosis oligonucleotides are included (left side). (B) The five classes of proteases are logically arranged together on the microarray. Thus, the microarray contains oligonucleotides for 21 aspartic proteases, 148 cysteine proteases, 184 metallo proteases, 177 serine proteases, 28 threonine proteases and 157 inhibitors as well as a further 46 positive and 7 negative controls (light areas of the array) spotted in triplicate. (C) Representative expression pattern of proteases and inhibitors in human mammary epithelial cells (cyanine 3 channel). The color bar shows the expression level: black no expression to white high expression. (1) (2) CLIP-CHIP • Oligonucelotides (571) for each protease, inactive homologue and inhibitor were obtained from the Human Genome Oligo Set v2.0 (Qiagen). Oligonucleotides were synthesized as 70-mers with a 5’- C6 amino linker for attachment to an epoxy silane glass matrix. • Oligonucleotides for the 144 genes not represented in the Human Oligo Set were synthesized after extensive BLAST analyses to identify unique sequences. • 715 protease and inhibitor oligonucleotides plus 46 positive and 7 negative control oligonucleotides were arranged in logical groups and spotted in triplicate (Figure 1). • A dedicated subarray is customized according to the experiment. Here we used 217 oligonucleotides for breast cancer related and prognosis genes as well as 192 control oligonucleotides (96 positive, 80 negative, 16 Array Controls/Ambion). Figure 3. Signal Intensities of proteases and inhibitors highly expressed in normal human mammary epithelial cells (Normal 1, 2) and invasive ductual breast carcinoma cells (Cancer 1,2) are listed. Normal1/Cancer1 and Normal2/Cancer2 were hybridized to the same slide. Normal mammary epithelial cells is from one source (BioCain); Cancer 1 and 2 are from two different patients with invasive ductual carcinoma Protease/Inhibitor No. Name Normal1 Normal2 Cancer 1Cancer 2 Aspartic A2 Pepsin C 64000 27000 64756 25460 A6 Cathepsin D 65000 27500 65002 29120 Cysteine C31 Ubiquitin C-term. Hydrolase BAP1 20077 22235 5916 C43 Caspase-8 21121 23723 6300 C76 USP22 35216 31388 34739 20559 C93 USP40 36904 18424 32863 8600 C122 Sentrin/SUMO protease 8 32850 39156 30488 21646 C142 Cezanne-2 22236 21060 Metallo M42 MMP28 9392 10307 17253 14200 M67 ADAM30 12098 38877 21658 M76 ADAMTS7 16048 54231 18457 31975 M85 ADAMTS17 10095 32566 19326 28476 M100 Carboxypeptidase A5 17783 37436 34066 29186 M107 Carboxypeptidase M 15853 9540 30530 18800 M117 Mitochondrial processing protease 10517 30183 13600 M121 Aminopeptidase like 1 12880 48155 28981 M126 Glu-carboxypeptidase-like 1 45817 65298 65039 65084 M134 Methionyl-aminopeptidase like 1 13914 32079 22734 19003 Serine S4 Kallikrein hK4 27039 31006 42279 S31 Tryptase gamma 16359 31245 46262 S70 Matriptase-2 21665 45452 52752 59200 S085 Pancreatic endopeptidase E (B)25081 34107 48282 S100 Plasminogen 43514 65448 30252 65535 Threonine T4 Proteosome catalytic subunit 1i 10302 3366 T11 Proteasome beta 4 subunit 34369 8355 10909 23147 T23 Gamma-glutamyltransferase 1 65440 42873 32568 65535 T25 Gamma-glutamyltransferase 2 6634 Inhibitors I60 Protease Inhibitor 9 / CAP3 23623 I69 Yukopin (SerpinB12) 23387 62902 20377 24169 I71 Antithrombin III 20259 37993 13389 22475 I86 Antileukoproteinase 18396 31798 18696 21702 I98 WAP four-disulfide core like 122167 43752 26509 18740 I102 Cystatin SA 23451 40947 I122 IGF binding protein 2 11668 35163 24800 28026 I142 Pregnancy zone protein 13788 48198 19918 18973 I152 Histatin 3 26400 33642 Figure 2. Expression levels of the 23 MMPs in normal human mammary epithelia and breast cancer cells. Matrix metalloproteinases (MMPs) are an important family of proteases that correlate with tumor grade. MMPs traditionally were thought to degrade the ECM in metastasis, but the large number of bioactive mediators that are precisely processed by MMPs reveals that MMPs are important signaling proteases and so may explain the side effects of MMPI trials. 0 2000 4000 6000 8000 10000 12000 14000 16000 Normal Breast (Cy Breast Cancer (Cy
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Transcript of The human protease CLIP-CHIP: Genomic analysis of all 715 human protease and inhibitor gene...
The human protease CLIP-CHIP: Genomic analysis of all 715 human protease and inhibitor gene transcripts in human breast carcinoma
Reinhild Kappelhoff,Tom Ewart+, Xose S. Puente*, Arun Seth+, Carlos López-Otín* and Christopher M. Overall†
Departments of Biochemistry and Molecular Biology†, Oral Biological and Medical Sciences, University of British Columbia, Vancouver, BC, Canada ([email protected]); +Department of Molecular Pathology, Sunnybrook & Women's College Health Sciences Centre, Toronto, ON, Canada, *Departamento de
Bioquimica y Biologia Molecular, Universidad de Oviedo, Oviedo, Spain
Introduction• The availability of the human genome sequence has enabled the identification the degradome—the complete repertoire of proteases produced in man. By using primary information retrieved from public and private sequencing projects, together with data from the MEROPS, InterPro and Ensembl databases we have annotated a total of 558 genes encoding proteases or protease-homologs in the human genome and 157 inhibitors (Xose S. Puente, Luis M. Sanchez, Christopher M. Overall, and Carlos Lopez-Otin, Nature Rev Genetics 4, 544-558, 2003). • To date there are no complete protease microarray chips reported.
Hybridization, scanning and analyzing the CLIP-CHIP• Hybridization of labeled cDNA to the microarray was at 25C for 16 h in 5x SSC, 0.3 % SDS. Washing steps were carried out at 42C.• The CLIP-CHIP was scanned in an Affymetrix 428 (MWG) microarray laser scanner using 532 nm for the cyanine 3 channel and 635 nm for the cyanine 5 channel. Photo multiplier gains were set individually for each channel.
Conclusions and outlook• The development of the CLIP-CHIP represents the first complete dedicated human protease and inhibitor chip reported to date. • The CLIP-CHIP will prove to be an extremely valuable and versatile tool for the genomic analysis of proteolytic function in vivo. • The advantage of using a custom glass chip is that new protease or inhibitor oligonucleotides can be added in an agile manner at each chip printing for minimal cost. • Moreover, having a dedicated subarray containing oligonucleotides relevant to the experimental question being addressed further focuses the experiment without distraction from analyses of many thousands of genes that is possible in larger arrays or the multiple 20mer oligonucleotides per gene necessary for the Affymetrix chips. • Work is in progress to analyze RNA from an additional 50 breast cancer tissues of different disease stages to analyze their protease and inhibitor expression profiles.
The CLIP-CHIP A dedicated human protease chip
CLIP
CH
IP
A1 A2 A3 A4 A5 K1 K2 A6 A7 A8 A9 A10 A11 A12 A13 A14 A15 N1 A16 A17 A18 A19 A20 A21
C1 C2 C3 C4 C5 K3 K4 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 K1 K2 C16 C17 C18 C19 C20
C21 C22 C23 C24 C25 C26 C27 C28 C29 C30 C31 C32 C33 C34 C35 C36 C37 K3 K4 C38 C39 C40 C41 C42
C43 C44 C45 C46 C47 C48 C49 C50 C51 C52 C53 N2 N3 C54 C55 C56 C57 C58 C59 C60 C61 C62 C63 C64
C65 C66 C67 C68 C69 C70 C71 C72 C73 C74 C75 N4 N5 C76 C77 C78 C79 C80 C81 C82 C83 C84 C85 C86
C87 C88 C89 C90 C91 C92 C93 C94 C95 C96 C97 C98 C99 C100 C101 C102 C103 C104 C105 C106 C107 C108 C109 C110
C111 C112 C113 C114 C115 K5 K6 C116 C117 C118 C119 C120 C121 C122 C123 C124 C125 K5 K6 C126 C127 C128 C129 C130
N6 C131 C132 C133 C134 K7 K8 C135 C136 C137 C138 C139 C140 C141 C142 C143 C144 K7 K8 C145 C146 C147 C148 N7
M1 M2 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 M13 M14 M15 M16 M17 M18 M19 M20 M21 M22 M23 M24
M25 M26 M27 M28 M29 M30 M31 M32 M33 M34 M35 M36 M37 M38 M39 M40 M41 M42 M43 M44 M45 M46 M47 M48
M49 M50 M51 M52 M53 M54 M55 M56 M57 M58 M59 M60 M61 M62 M63 M64 M65 M66 M67 M68 M69 M70 M71 M72
M73 M74 M75 M76 M77 K12 K9 M78 M79 M80 M81 M82 M83 M84 M85 M86 M87 K12 K9 M88 M89 M90 M91 M92
M93 M94 M95 M96 M97 K10 K11 M98 M99 M100 M101 M102 M103 M104 M105 M106 M107 K10 K11 M108 M109 M110 M111 M112
M113 M114 M115 M116 M117 M118 M119 M120 M121 M122 M123 M124 M125 M126 M127 M128 M129 M130 M131 M132 M133 M134 M135 M136
M137 M138 M139 M140 M141 M142 M143 M144 M145 M146 M147 M148 M149 M150 M151 M152 M153 M154 M155 M156 M157 M158 M159 M160
M161 M162 M163 M164 M165 M166 M167 M168 M169 M170 M171 M172 M173 M174 M175 M176 M177 M178 M179 M180 M181 M182 M183 M184
S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12 S13 S14 S15 S16 S17 S18 S19 S20 S21 S22 S23 S24
S25 S26 S27 S28 S29 S30 S31 S32 S33 S34 S35 S36 S37 S38 S39 S40 S41 S42 S43 S44 S45 S46 S47 S48
S49 S50 S51 S52 S53 S54 S55 S56 S57 S58 S59 S60 S61 S62 S63 S64 S65 S66 S67 S68 S69 S70 S71 S72
S73 S74 S75 S76 S77 K1 K2 S78 S79 S80 S81 S82 S83 S84 S85 S86 S87 K1 K2 S88 S89 S90 S91 S92
S93 S94 S95 S96 S97 K3 K4 S98 S99 S100 S101 S102 S103 S104 S105 S106 S107 K3 K4 S108 S109 S110 S111 S112
S113 S114 S115 S116 S117 S118 S119 S120 S121 S122 S123 S124 S125 S126 S127 S128 S129 S130 S131 S132 S133 S134 S135 S136
S137 S138 S139 S140 S141 S142 S143 S144 S145 S146 S147 S148 S149 S150 S151 S152 S153 S154 S155 S156 S157 S158 S159 S160
S161 S162 S163 S164 S165 S166 S167 S168 S169 S170 S171 S172 S173 S174 S175 S176 S177 K5 K6 K7 T1 T2 T3 T4
T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16 T17 T18 T19 T20 T21 T22 T23 T24 T25 T26 T27 T28
I1 I2 I3 I4 I5 I6 I7 I8 I9 I10 I11 I12 I13 I14 I15 I16 I17 I18 I19 I20 I21 I22 I23 I24
I25 I26 I27 I28 I29 I30 I31 I32 I33 I34 I35 I36 I37 I38 I39 I40 I41 I42 I43 I44 I45 I46 I47 I48
I49 I50 I51 I52 I53 K12 K9 I54 I55 I56 I57 I58 I59 I60 I61 I62 I63 K12 K9 I64 I65 I66 I67 I68
I69 I70 I71 I72 I73 K10 K11 I74 I75 I76 I77 I78 I79 I80 I81 I82 I83 K10 K11 I84 I85 I86 I87 I88
I89 I90 I91 I92 I93 I94 I95 I96 I97 I98 I99 I100 I101 I102 I103 I104 I105 I106 I107 I108 I109 I110 I111 I112
I113 I114 I115 I116 I117 I118 I119 I120 I121 I122 I123 I124 I125 I126 I127 I128 I129 I130 I131 I132 I133 I134 I135 I136
I137 I138 I139 I140 I141 I142 I143 I144 I145 I146 I147 I148 I149 I150 I151 I152 I153 I154 I155 I156 I157 K5 K6 K7
Serine (177)
Threonine (28)
Inhibitors(157)
Aspartic(21)
Cysteine(148)
Metallo (184)
(+) Controls (46)
(-) Controls (7)
(A)
(B)
Inhibitors
SerineCysteine
Metallo
Aspartate
Threonine
(C)
low expression level high
Figure 1. The human protease CLIP-CHIP. (A) Overview of the CLIP-CHIP microarray layout. The human microarray is spotted in duplicate (1, 2). In addition 192 control oligonuclotides and a dedicated subarray with 217 breast cancer related and prognosis oligonucleotides are included (left side). (B) The five classes of proteases are logically arranged together on the microarray. Thus, the microarray contains oligonucleotides for 21 aspartic proteases, 148 cysteine proteases, 184 metallo proteases, 177 serine proteases, 28 threonine proteases and 157 inhibitors as well as a further 46 positive and 7 negative controls (light areas of the array) spotted in triplicate. (C) Representative expression pattern of proteases and inhibitors in human mammary epithelial cells (cyanine 3 channel). The color bar shows the expression level: black no expression to white high expression.
(1) (2)
CLIP-CHIP• Oligonucelotides (571) for each protease, inactive homologue and inhibitor were obtained from the Human Genome Oligo Set v2.0 (Qiagen). Oligonucleotides were synthesized as 70-mers with a 5’-C6 amino linker for attachment to an epoxy silane glass matrix. • Oligonucleotides for the 144 genes not represented in the Human Oligo Set were synthesized after extensive BLAST analyses to identify unique sequences. • 715 protease and inhibitor oligonucleotides plus 46 positive and 7 negative control oligonucleotides were arranged in logical groups and spotted in triplicate (Figure 1).• A dedicated subarray is customized according to the experiment. Here we used 217 oligonucleotides for breast cancer related and prognosis genes as well as 192 control oligonucleotides (96 positive, 80 negative, 16 Array Controls/Ambion).
Figure 3. Signal Intensities of proteases and inhibitors highly expressed in normal human mammary epithelial cells (Normal 1, 2) and invasive ductual breast carcinoma cells (Cancer 1,2) are listed. Normal1/Cancer1 and Normal2/Cancer2 were hybridized to the same slide. Normal mammary epithelial cells is from one source (BioCain); Cancer 1 and 2 are from two different patients with invasive ductual carcinoma
Protease/InhibitorNo. Name Normal1Normal2Cancer 1Cancer 2Aspartic A2 Pepsin C 64000 27000 64756 25460
A6 Cathepsin D 65000 27500 65002 29120Cysteine C31 Ubiquitin C-term. Hydrolase BAP1 20077 22235 5916
C43 Caspase-8 21121 23723 6300C76 USP22 35216 31388 34739 20559C93 USP40 36904 18424 32863 8600C122 Sentrin/SUMO protease 8 32850 39156 30488 21646C142 Cezanne-2 22236 21060
Metallo M42 MMP28 9392 10307 17253 14200M67 ADAM30 12098 38877 21658M76 ADAMTS7 16048 54231 18457 31975M85 ADAMTS17 10095 32566 19326 28476M100 Carboxypeptidase A5 17783 37436 34066 29186M107 Carboxypeptidase M 15853 9540 30530 18800M117Mitochondrial processing protease 10517 30183 13600M121Aminopeptidase like 1 12880 48155 28981M126 Glu-carboxypeptidase-like 1 45817 65298 65039 65084M134 Methionyl-aminopeptidase like 1 13914 32079 22734 19003
Serine S4 Kallikrein hK4 27039 31006 42279S31 Tryptase gamma 16359 31245 46262S70 Matriptase-2 21665 45452 52752 59200S085 Pancreatic endopeptidase E (B) 25081 34107 48282S100 Plasminogen 43514 65448 30252 65535
Threonine T4 Proteosome catalytic subunit 1i 10302 3366T11 Proteasome beta 4 subunit 34369 8355 10909 23147T23 Gamma-glutamyltransferase 1 65440 42873 32568 65535T25 Gamma-glutamyltransferase 2 6634
Inhibitors I60 Protease Inhibitor 9 / CAP3 23623I69 Yukopin (SerpinB12) 23387 62902 20377 24169I71 Antithrombin III 20259 37993 13389 22475I86 Antileukoproteinase 18396 31798 18696 21702I98 WAP four-disulfide core like 1 22167 43752 26509 18740I102 Cystatin SA 23451 40947I122 IGF binding protein 2 11668 35163 24800 28026I142 Pregnancy zone protein 13788 48198 19918 18973I152 Histatin 3 26400 33642
Figure 2. Expression levels of the 23 MMPs in normal human mammary epithelia and breast cancer cells.Matrix metalloproteinases (MMPs) are an important family of proteases that correlate with tumor grade. MMPs traditionally were thought to degrade the ECM in metastasis, but the large number of bioactive mediators that are precisely processed by MMPs reveals that MMPs are important signaling proteases and so may explain the side effects of MMPI trials.
0
2000
4000
6000
8000
10000
12000
14000
16000
MMP 1MMP 2MMP 3MMP 7MMP 8MMP 9MMP10MMP11MMP12MMP13MMP14MMP15MMP16MMP17MMP19MMP20MMP21MMP23MMP24MMP25MMP26MMP27MMP28
Signal Intensity
Normal Breast (Cy3)
Breast Cancer (Cy5)
![Protease Inhibitors - DNA GDAŃSK€¦ · Tab. 1: Individual Protease Inhibitors Prod.-No. Description M [g/mol] Structure Target Protease Class, Target Enzymes Mechanism Recommen-ded](https://static.fdocuments.in/doc/165x107/5ad6ea197f8b9a5b538bf718/protease-inhibitors-dna-gdansk-tab-1-individual-protease-inhibitors-prod-no.jpg)
