Abstract
Human germinal center (GC)-associated lymphoma (HGAL) is an adaptor protein expressed in GC B cells. HGAL regulates cell motility and B-cell receptor (BCR) signaling, processes that are central for the successful completion of the GC reaction. Herein, we demonstrate phosphorylation of HGAL by Syk and Lyn kinases at tyrosines Y80, Y86, Y106Y107, Y128, and Y148. The HGAL YEN motif (amino acids 107-109) is similar to the phosphopeptide motif pYXN used as a binding site to the growth factor receptor-bound protein 2 (Grb2). We demonstrate by biochemical and molecular methodologies that HGAL directly interacts with Grb2. Concordantly, microscopy studies demonstrate HGAL-Grb2 colocalization in the membrane central supramolecular activation clusters (cSMAC) following BCR activation. Mutation of the HGAL putative binding site to Grb2 abrogates the interaction between these proteins. Further, this HGAL mutant localizes exclusively in the peripheral SMAC and decreases the rate and intensity of BCR accumulation in the cSMAC. Furthermore, we demonstrate that Grb2, HGAL, and Syk interact in the same complex, but Grb2 does not modulate the effects of HGAL on Syk kinase activity. Overall, the interplay between the HGAL and Grb2 regulates the magnitude of BCR signaling and synapse formation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2286-2297 |
| Number of pages | 12 |
| Journal | Blood Advances |
| Volume | 3 |
| Issue number | 15 |
| DOIs | |
| State | Published - Aug 13 2019 |
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ASJC Scopus subject areas
- Medicine(all)
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Interplay between HGAL and Grb2 proteins regulates B-cell receptor signaling. / Jiang, Xiaoyu; Lu, Xiaoqing; Zhang, Yu; Lacaria, Leda; Schuchardt, Brett J.; Mikles, David C.; Magistri, Marco; García-Ramírez, Idoia; Sanchez-Garcia, Isidro; Farooq, Amjad; Verdun, Ramiro E.; Abdulreda, Midhat H.; Moy, Vincent T.; Lossos, Izidore S.
In: Blood Advances, Vol. 3, No. 15, 13.08.2019, p. 2286-2297.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interplay between HGAL and Grb2 proteins regulates B-cell receptor signaling
AU - Jiang, Xiaoyu
AU - Lu, Xiaoqing
AU - Zhang, Yu
AU - Lacaria, Leda
AU - Schuchardt, Brett J.
AU - Mikles, David C.
AU - Magistri, Marco
AU - García-Ramírez, Idoia
AU - Sanchez-Garcia, Isidro
AU - Farooq, Amjad
AU - Verdun, Ramiro E.
AU - Abdulreda, Midhat H.
AU - Moy, Vincent T.
AU - Lossos, Izidore S.
PY - 2019/8/13
Y1 - 2019/8/13
N2 - Human germinal center (GC)-associated lymphoma (HGAL) is an adaptor protein expressed in GC B cells. HGAL regulates cell motility and B-cell receptor (BCR) signaling, processes that are central for the successful completion of the GC reaction. Herein, we demonstrate phosphorylation of HGAL by Syk and Lyn kinases at tyrosines Y80, Y86, Y106Y107, Y128, and Y148. The HGAL YEN motif (amino acids 107-109) is similar to the phosphopeptide motif pYXN used as a binding site to the growth factor receptor-bound protein 2 (Grb2). We demonstrate by biochemical and molecular methodologies that HGAL directly interacts with Grb2. Concordantly, microscopy studies demonstrate HGAL-Grb2 colocalization in the membrane central supramolecular activation clusters (cSMAC) following BCR activation. Mutation of the HGAL putative binding site to Grb2 abrogates the interaction between these proteins. Further, this HGAL mutant localizes exclusively in the peripheral SMAC and decreases the rate and intensity of BCR accumulation in the cSMAC. Furthermore, we demonstrate that Grb2, HGAL, and Syk interact in the same complex, but Grb2 does not modulate the effects of HGAL on Syk kinase activity. Overall, the interplay between the HGAL and Grb2 regulates the magnitude of BCR signaling and synapse formation.
AB - Human germinal center (GC)-associated lymphoma (HGAL) is an adaptor protein expressed in GC B cells. HGAL regulates cell motility and B-cell receptor (BCR) signaling, processes that are central for the successful completion of the GC reaction. Herein, we demonstrate phosphorylation of HGAL by Syk and Lyn kinases at tyrosines Y80, Y86, Y106Y107, Y128, and Y148. The HGAL YEN motif (amino acids 107-109) is similar to the phosphopeptide motif pYXN used as a binding site to the growth factor receptor-bound protein 2 (Grb2). We demonstrate by biochemical and molecular methodologies that HGAL directly interacts with Grb2. Concordantly, microscopy studies demonstrate HGAL-Grb2 colocalization in the membrane central supramolecular activation clusters (cSMAC) following BCR activation. Mutation of the HGAL putative binding site to Grb2 abrogates the interaction between these proteins. Further, this HGAL mutant localizes exclusively in the peripheral SMAC and decreases the rate and intensity of BCR accumulation in the cSMAC. Furthermore, we demonstrate that Grb2, HGAL, and Syk interact in the same complex, but Grb2 does not modulate the effects of HGAL on Syk kinase activity. Overall, the interplay between the HGAL and Grb2 regulates the magnitude of BCR signaling and synapse formation.
UR - http://www.scopus.com/inward/record.url?scp=85070683848&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85070683848&partnerID=8YFLogxK
U2 - 10.1182/bloodadvances.2018016162
DO - 10.1182/bloodadvances.2018016162
M3 - Article
C2 - 31362927
AN - SCOPUS:85070683848
VL - 3
SP - 2286
EP - 2297
JO - Blood advances
JF - Blood advances
SN - 2473-9529
IS - 15
ER -